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Evolutionary conservation of protein interaction properties has been shown to be a valuable indication for functional importance. Here we use homology interface modeling of 10 Ras-effector complexes by selecting ortholog proteins from 12 organisms representing the major eukaryotic branches, except plants. We find that with increasing divergence time the sequence similarity decreases with respect to the human protein, but the affinities and association rate constants are conserved as predicted by the protein design algorithm, FoldX. In parallel we have done computer simulations on a minimal network based on Ras-effector interactions, and our results indicate that in the absence of negative feedback, changes in kinetics that result in similar binding constants have strong consequences on network behavior. This, together with the previous results, suggests an important biological role, not only for equilibrium binding constants but also for kinetics in signaling processes involving Ras-effector interactions. Our findings are important to take into consideration in system biology approaches and simulations of biological networks.
Author Summary
Cellular signal transductions processes are based on protein interactions. Proteins can either associate transiently with each other or form stable complexes, and the strength of the interaction is described by the affinity (the affinity is the ratio between the rate of dissociation and association). Protein complexes with similar affinities can bind and dissociate with different rates, and these rates describe the kinetic properties of protein binding. These kinetic rates are important for signaling; however, to what extent individual changes in such rate constants are biologically important or whether the affinity is more crucial might be different in different signaling processes. In this study we analyze whether association rates are conserved during evolution, because evolutionary conservation of protein biochemical properties is usually a valuable indication of its importance. We analyzed the binding of Ras proteins to effector domains, which are central proteins in many signal transduction pathways, in different organisms. On the basis of homology modeling and energy calculations we find that association rates are conserved, although the sequence similarity decreases compared to the human protein. Our finding should encourage further analysis of the importance of kinetics for cellular signal transduction.
Citation: Kiel C, Aydin D, Serrano L (2008) Association Rate Constants of Ras-Effector Interactions Are Evolutionarily Conserved. PLoS Comput Biol 4(12): e1000245. doi:10.1371/journal.pcbi.1000245
Editor: Christopher Rao, University of Illinois at Urbana-Champaign, United States of America
Received: July 7, 2008; Accepted: November 6, 2008; Published: December 19, 2008
Copyright: © 2008 Kiel et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by EU INTERACTION PROTEOME (Grant No. LSHG-CT-2003-505520).
Competing interests: The authors have declared that no competing interests exist.
Protein-protein interactions are the central elements in all signal transduction processes. The life times of protein complexes as well as regulatory processes need to be tightly controlled for proper systems functioning. Affinities are used to characterize the strength of protein interactions and the affinities between proteins involved in signaling processes have been shown to correlate with the activities (output/response) in such signal transduction processes [1],[2]. In the majority of the cases, affinities between proteins and protein-ligands are determined using equilibrium binding methods, like isothermal titration calorimetry and fluorescence based methods, while rate constants of association and dissociation are only rarely determined. However, correlations of either association or dissociation rate constants with in vivo activity suggest that kinetic properties play a role in the cellular context [3]–[7]. As the affinity (Kd) can be described as the ratio between the dissociation (koff) and association (kon) rate constants, different ratios of kon and koff values can give rise to similar affinities. Kinetic rate constants have been shown to be important for signal transduction, however to which extent kinetics influence signaling might depend on the actual network and network topology. We could speculate that fast kon and koff values could result in rapid activation and deactivation upon short pulses of a stimulus, while slow ones could filter noise and result in prolonged signaling. If this is true it might open new aspects of cellular signal transduction regulation and could probably lead to conceptually new strategies in drug design. It is likely that the answer will depend on the network topology: rate constants might be important in some signaling branches, in others not.
Evolutionary conservation of protein composition and biochemical properties is usually a valuable indication for the cellular importance of a specific protein complex. In this study we have selected the Ras-effector complex formation, in order to analyze whether kinetic rate constants are evolutionary conserved. Ras proteins belong to the Ras superfamily of small GTPases and they have key roles in various signal transduction pathways, like proliferation and differentiation [8]. They act as molecular switches by cycling between an active GTP-bound and an inactive GDP-bound state [9],[10]. Active Ras (Ras·GTP) can interact with effector molecules such as the Ser/Thr kinase Raf. The resulting Raf activation triggers the MAP kinase pathway, which leads to the transcription of target genes in the nucleus [11],[12]. Other Ras·GTP binding effector proteins that have been identified are the PI3-kinase, members of the RalGDS family, and AF6 [13]–[16]. Effector proteins bind to Ras·GTP via a common domain with a ubiquitin-like topology [17]–[22], and various structures of effector domains in complex with Ras proteins have revealed a similar binding mode that involves mainly two antiparallel ß-sheets of the RBD and Ras, respectively [23]–[29].
As Ras-effector protein interactions play a key role in cells, pathways involving Ras-effector interactions can be assumed to be at least partially conserved during evolution. In this study we analyzed whether the affinities and the association rate constants are conserved for 10 Ras-effector complexes in 12 different species, including worms, flies, fishes, and mammalian organisms. We used homology interface modeling and energy calculations, using FoldX 2.8 (http://foldx.crg.es/) [30],[31] in order to model Ras-effector interactions of proteins from different organisms. FoldX uses an algorithm based on the original work of Schreiber and co-workers to calculate relative changes in kon which has been validated experimentally numerous times [32]. Homology modeling was performed in a similar way as done in a previous study, on a genome-wide level for all human Ras-effecter complexes [33],[34].
Importance of Electrostatic Charge Complementarity for Ras-Effector Association Kinetics
Binding of effector proteins to Ras proteins is mediated via a domain with an ubiquitin-like topology [35]. Members of the ubiquitin domain superfamily are the RA, the RBD, the PI3Krbd, the UBQ and the B41/ERM domain families [36]. However, the binding of Ras to effector domains does not depend on the fold itself, but rather on certain amino acid residues on the surface that are crucial for binding.
An important observation found in Ras-effector complex structures is the high charge complementarity between the proteins of the complex, where Ras is mainly negatively charged and the effector RBDs are mainly positively charged [23]–[29]. Various studies have shown that a strong electrostatic surface complementarity in a protein complex enhances the association rate constant by forming of a low affinity encounter complex before the final high affinity complex is formed [37]–[42]. The complex formation itself is promoted by electrostatic steering which stabilizes the transition state by decreasing the energy barrier for association [42],[43]. In agreement with this concept of electrostatic steering and encounter complex formation, the association rate constants between Ras and effector domains were found to be fast (reviewed in [44]). Interestingly, the variance in binding energies when comparing different Ras-effector complexes is mainly the consequence of different association rate constants, while the dissociation rate constants are in a similar range [45]–[47]. For example, RafRBD is highly positively charged in its Ras binding region, and here the association rate constant was found to be very high in complex with the mainly negatively charged Ras proteins. In contrast, RalGDS has a mixed charged distribution (Figure 1A), and the kon in complex with Ras is much lower. Interestingly, introducing positively charged residues at the edge of the interface of RalGDS can change binding kinetics and these RalGDS mutants were shown to bind "Raf-like" to Ras [43]. In Figure 1B–F we show the electrostatic surface potentials of several other RA/RB domains, which can bind to Ras (Rgl1, Rgl2, Grb7, AF6_RA1, PLCe_RA2) and for which structures have been solved, either by NMR or X-Ray, and we orient them similar as the RA domain of RalGDS in complex with Ras. In all cases the interface surface areas have a strong positively charged electrostatic potential, which suggests that association kinetics are important for these RA/RBD domains as well.
(A) The Ras-RalGDS complex (pdb entry: 1lfd), and they single domains of (B) Rgl1 (pdb-entry: 1ef5), (C) Rgl2 (pdb-entry: 1rlf), (D) Grb7 (pdb-entry: 1wgr), (E) AF6-RA1 (pdb-entry: 1wxa), and (F) PLCeRA2 (pdb-entry: 2c5l) are shown.
FoldX Electrostatic Interactions and Association Rate Constants Correlate with Experimental Association Rate Constants in Ras/Raf and Ras/RalGDS Complexes
Although the algorithm developed by Schreiber and co-workers implemented in FoldX (http://foldx.crg.es/) [30],[31], has been validated experimentally on many different proteins, still it is a prediction method and as such needs some validation on the particular system under study. For this, we have selected the Ras-Raf complex and calculated kon values (ΔG kon) at different salt concentrations, ranging from 0 to 800 mM NaCl (corresponding to an ionic strength of ∼50 to 850 mM in 50 mM Tris-buffer), and compared these results with experimental kon values measured at different ionic strength using stopped-flow (Table S1; [48]). The experimental kon values range from 7.4 to 60 µM−1 s−1 and an excellent correlation with calculated association rate constants was observed (R = 0.99) (Figure 2A). Further, we used FoldX in order to generate in silico a series of mutations of charged residue in RalGDS, located either in the binding site, or at the edge of the binding site, and we calculated binding energies as well as association rate constants using the Ras-Ral complex. When comparing these results with experiments [43], we find again a very good correlation between experimental and calculated kon values (Figure 2B) (R = 0.89), with the slopes of the two correlations (ionic strength and mutants) been similar. This indicates that absolute values of association rate constants can be reliably calculated over a wide range for different ionic strengths and mutations of Ras-effector complexes.
(A) Ras-Raf complex at different ionic strength (salt concentrations form 0 to 800 mM NaCl) (Table S1, [48]). The correlation coefficient is 0.987. (B) The Ras-RalGDS complex. Mutations within the RalGDS domain were introduced, and compared with experimental data (Table S1, [43]). The correlation coefficient is 0.892.
Ortholog Prediction of Ras and Selected Effector Proteins
We have selected proteins containing RA, RBD, PI3Krbd, and B41 domains, similar as in our previous genome-wide Ras-effector homology interface modeling study [34] (Figure 3A), for which binding to Ras has been shown experimentally (Table 1). These include the different isoforms of the Raf kinases, RalGDS, and the related proteins, Rgl1, and Rgl2. Other Ras binding domains are the PI3K-p110 gamma subunit, and Krit. In the following we will often refer to members of the ubiquitin superfamily as UBDs, without differentiating between RA, RBD, PI3Krbd, or B41.
(A) Domain architecture of selected effector proteins. The ubiquitin-like domains mediating binding to Ras (RA, RBD, PI3K_rbd, and B41) are shown in yellow, other domains in grey. The domain prediction was done using SMART [52],[53] using the respective sequence from Homo sapiens. (B) The relationship and divergence time (million of years ago (Mya)±standard error) of used model organisms are shown. The divergence times for the chordate – arthropod divergence and the one for divergence of nematodes from the lineage leading to chordates and arthropods were derived from Wang et al 1999 [61]. The others were taken from Hedges 2002 [62].
In order to derive Ras and effector protein "interactions" from organisms representing the major eukaryotic branches, we have selected the following species (Figure 3B): Homo sapiens (hs) and Mus musculus (mm) were chosen for mammals, Gallus gallus (gg) for birds, Xenopus tropicalis (xt) for amphibians, Fugu rubripes (fr), and Dario rerio (dr) for fishes, Drosophila melanogaster (dm), Drosophila pseudoobscura (dp), Anopheles gambiae (ag) and Apis mellifera (am) for arthropods, and Caenorhabditis elegans (ce) and Caenorhabditis briggsae (cb) for nematodes. The orthologs were predicted by using the ENSEMBL (http://www.ensembl.org) [49] and the IMPARANOID databases (http://inparanoid.cgb.ki.se/) [50],[51]. Domains were predicted using SMART [52],[53] and the sequences were aligned automatically and by manual curation taking structural information into account [35] (Figure S1). Depending on the organism, between 22% and 78% of all human proteins orthologs were identified. When taking into account that certain proteins in lower organisms are orthologs of more than one human protein, e.g., RalGDS of C. elegans is also an ortholog of Rgl1 and Rgl2, the number of orthologs in different organisms ranges from 33% to 100%. The alignments of the UBDs of Ras effector proteins show a high similarity within orthologs and often also between different proteins of the same domain family. Furthermore, the similarity within the secondary structures of the RBD is higher than within the loops, indicating a conservation of the binding mode. The sequence identity of ortholog proteins (for detailed description see method) ranges between 100 and ∼20% (Table S2). However, in the majority of the cases the sequence identity decreases to ∼30/40%. The only exceptions are the different PI3kinase p110 isoforms, where a drastic drop in sequence identity is observed for the corresponding othologs/isoforms in C. elegans/C. briggsae.
The sequences of proteins that have a key role in cells are usually highly conserved among all organisms. In accordance with this, the sequences of Ras proteins were found to be nearly identical, especially in the effector binding region (Figure S2). The three Ras proteins, H-Ras, N-Ras and K-Ras could only be found in vertebrates, for arthropods and nematodes there is only one Ras protein which is most likely to be an ortholog of H-Ras. Due to the similarity in the effector binding region, only HRas was modeled (here termed as Ras).
Homology Interface Modeling of Ortholog Ras-Effector Complexes
The first three secondary structure elements (β1, β2, and α1) of the ubiquitin-like domain determine the interaction surface towards Ras and they have the largest impact on binding energy of the complex [33]. In those cases in which a crystal structure of Ras in complex with a RBD domain was available we use the structure to model the ortholog sequences (Ras-Raf, Ras-Ral, Ras-PI3 Kinase, Ras-Byr). For the rest we used the templates modeled in our previous study [34] (Table S3) that were validated experimentally by pull-down experiments (for details see methods). Only those UB domains that could be reliably modeled were selected (e.g., no van der waals' clashes above a fixed threshold of 2 kcal/mol). The species were then grouped into human (hs), mouse (mm), birds (gg), amphibians (xt), insects (dm, dp, ag, am) and nematodes (ce, cb). The mean of ΔG and ΔGkon within each group was calculated and taken as value for the complete group. By grouping the different organisms, the problem of missing sequences can be solved for many proteins and mean values as well as standard deviation of ΔG and ΔGkon can be calculated (the results do not change if we consider individual organisms, data not shown).
The results for all interaction energies (ΔGint FoldX) and contribution of association rate constants (ΔGkon FoldX) were plotted against the divergence time (Table S4 and Figure S3). While the sequence similarity decreases with increasing divergence time, the interaction energies as well as the association rate constants are conserved. A selection of representative results is shown in Figure 4. A comparison of the mean values for all interaction energies and kon values calculated for a particular Ras-effector complex in different organisms shows that the standard deviations are in the majority of the complexes small (Table S5 and Figure 5A). Interestingly, the interaction energies correlate with the association rate constant contribution (Figure 5B) (R = 0.71). This indicates that also for the so far kinetically uncharacterized UBD domains, the changes in ΔG are mainly a consequence of changing kon. Thus, this could be underlying binding principle for the complete Ras-effector family.
Representative Results for predicted association rate constants. Association rate constants are conserved for Ras-effector complexes in different organisms with increasing divergence time, while the sequence identity is decreasing. Color code: AF6_RA2 (orange), RalGDS (dark blue), BRaf (light blue), CRaf (dark green), and AF6_RA1 (light green).
(A) Results from UBDs from the same protein or from the same protein family were shown in similar color. The results were taken from Table S5. (B) Correlation of calculated interaction energies with calculated association rate constants. The results were taken from Table S5, and the correlation coefficient is 0.71.
In order to demonstrate that large changes in association rate constants would have been possible theoretically, we have selected the human Ras-RalGDS complex as an example for an in silico mutagenesis using FoldX. By either introducing positively or negatively charged residues at all positions at the surface of RalGDS, the FoldX-kon contribution could be increased from −3.65 kcal/mol to −7,6 kcal/mol or decreased to −0.47 kcal/mol, respectively (data not shown).
Simulation of a Minimal Ras-Effector Network
In order to analyze whether compensating changes in kon and koff can influence signal transduction, we used in silico simulations of a sub-network within the EGF signal transduction pathway. Activation of proteins following EGF stimulation is one of the most studied signaling systems, which involves the Ras-CRaf interaction as central elements, and numerous simulation models exist, which are able to correctly predict different aspects of EFG signaling found experimentally [54]–[59]. Based on these earlier models we have constructed a minimal network involving Ras and Raf kinase (Figure 6A and Table S6). This minimal model involves activation of GEF upon stimulation (A), which results in activation of Ras (RasT = RasGTP). Subsequent binding of Raf to RasT activates Raf (Raf_act), which in turn leads to activation of a downstream target (X). Negative regulation was introduced by the GAP catalyzed hydrolysis reaction of RasGTP to RasGDP (RasD). We simulated this network by first applying a constant stimulus of "A" for 500 seconds using the wild type kon and koff values for the Ras-Raf interaction. Then we simulated the network with either 10-fold higher kon and koff rate constants, or 10 fold lower kon and koff (Figure 6B). Minor changes are observed when following X over time (activation peak) for the simulation with 10-fold higher kon and koff compared to the wild type situation. Only the simulation of 10-fold lower kon and koff resulted in a slightly smaller activation peak. However, when simulating the network by applying a pulse of stimulation, of 10 s of "A" (and then removing the stimulus), large changes in the activation peak are observed, with a higher maximum for the simulation of 10-fold higher kon and koff values for the Ras-Raf interaction (Figure 6C). This shows, that under certain cellular conditions, like short pulse of activation, large changes in activation are expected for mutants with similar affinity, but changed and compensating effects on kon and koff. Thus, kinetic properties can be crucial, and in the case of Ras-effector interactions, association kinetics will be important to result in sufficient activation, when the system is activated by applying a pulse.
(A) Schematic diagram of the reactions involved. For details see text and Table S5. Simulation of the network using the WT affinity of the Ras-Raf complex (blue line), 10 fold higher kon and koff rates (pink line), or 10 fold lower kon and koff values (green line). Either a constant stimulus was applied for 500 seconds (B) or the system was activated for 10 seconds and then the stimulus was removed (C).
The complex formation of Ras and effector proteins is driven by high association rate constants and only moderate dissociation rate constants [45]–[47]. Further, changes in affinity are mainly the consequence of changed association rate constants. Association rate constants can be influenced by mutating charged residues at the edge of the interface [32],[43]. If electrostatic interactions and association rate constants are important for the biological function of the cell, they should be conserved during the course of evolution. Using homology modeling and energy calculation covering a wide-range of sequences, and relating the output to the sequence conservation, we found that interaction energies as well as the electrostatic contributions and the association rate constants are conserved as well. While the sequence identity decreases with divergence time between the selected organisms, no trend could be found for the interaction energy and energies related to the electrostatics and kon, although theoretically it should be possible, when sampling the possible contributions of kon at different amino acid positions (Figure S4).
Biologically, electrostatic interactions within Ras-effector complex interfaces could be functionally important, because they are the basis for the observed dynamic behavior, as observed in the case of Ras binding to the Raf kinase effector protein: The Ras-RafRBD complex formation is characterized by both high association and dissociation rate constants (kon and koff), leading to affinities (Kd = koff/kon) in the range of 1 to 0.05 µM, under physiological conditions (this relatively low affinity seems to be functionally sufficient, since Ras is attached to the membrane via a lipid modification). The high kon values provide the possibility to have a fast dissociation of the complex, while still having a reasonable tight binding complex (the lifetime of the complex between Ras and RalGDS, for example, is 0.1 s-1; see reference [47]). As Ras signaling depends very crucially on a strict control through regulating proteins like GAPs (GTPase activating proteins) and GEFs (guanine nucleotide exchange factors), this fast dissociation allows regulatory proteins to access and act.
We assume that electrostatics contributions and binding kinetics could be important in other Ras signaling pathways, since association rate constants were found to be conserved during evolution, as demonstrated in this study for 10 effector domains. Further in vivo analysis will be needed to prove this hypothesis. These experiments could be performed by designing mutant variants, which are expected to have similar affinities, but changed association and dissociation rate constants. These protein variants could be expressed in cells and the effect on signal transduction monitored, e.g., after different pulses of stimulation.
It is expected that the effect of changing rate constants depends also on the network topology (negative feedback, feed forward inhibition, etc). This knowledge will be important for systems biology and simulation approaches, in order to know, at which positions in the network affinities will be sufficient, while for other accurate rate constants will be crucial for correct prediction. Further, it could open conceptually new aspects in drug design.
Selected Species, Orthologs, and Domain Prediction
Proteins from the following species were used in order to get a good representation of all branches: Homo sapiens and Mus musculus (mammals), Gallus gallus (birds), Xenopus tropicalis (amphibians), Fugu rubripes and Danio rerio (fishes), Drosophila melanogaster, Drosophila pseudoobscura, Anopheles gambiae and Apis mellifera (arthropods), and Caenorhabditis elegans and Caenorhabditis briggsae (nematodes). Only two RA domain containing were retrieved from Saccharomyces cerevisiae (sc), because these proteins are involved in a different pathway. For each protein, the human ENSEMBL protein ID was retrieved from ENSEMBL (http://www.ensembl.org) [49]. The orthologs were predicted by using the ENSEMBL database for Xenopus tropicalis, or the INPARANOID database (http://www.inparanoid.cgb.ki.se) [50],[51]. ENSEMBL [49] classifies the prediction based on the BLAST results. Only those orthologs were chosen that were a unique best reciprocal hit in both directions. As the INPARANOID database [50],[51] provides more information about orthologs of members of protein families, e.g., PI3K p110, the prediction was preferentially used. The sequences retrieved from ENSEMBL or INPARANOID were analyzed using SMART (http://smart.embl-heidelberg.de) [52],[53], in order to determine the domain architecture of the protein and the domain sequences.
Homology Modelling and Energy Calculations Using FoldX
For modeling of Ras-binding domains in complex with Ras proteins, we have taken the pdb-files of the following Ras effector complexes: Ras-RalGDS (pdb-entry: 1LFD), Ras-PI3Kinase (pdb-entry: 1HE8), and Raps-Raf (pdb-entry: 1GUA). Different template structures have been generated by deleting certain parts in the complex; the decision was mainly based on the alignment used to model the different binding domains. The ortholog sequences for one protein were aligned using standard automatic alignment tools, since sequence homology is high. However, the alignment of different effector domains from different families (RA, RBD, PI3Krbd, B41), was done based on manual curated structural-based sequence alignments as discussed in detail in a previous publication [35]. Basically two kinds of template structures have been generated (Table S3): a short version, where all secondary structure elements and loops (apart from ß1, ß1, ß2, a1) were deleted, as this is the part mainly contributing to the binding energy (similar as done in our previous study [33],[34]. In addition 'long template' structures have been generated. We could not model loop regions in those cases where the loops where not of the same length. For having a proline at the beginning of ß-strand 1 (position 26 in RalGDS, position 229 in PI3K, position 66 in Raf and position 81 in spByr2), we prepared special template structures by moving the backbone slightly, after introducing the proline at these positions (we checked that the proline was in acceptable dihedral angles and that the main chain CO group was still H-bonded to Ras). These template structures were then used to model the complex structures for AF6_RA2. The homology modelling was done as described before [33],[34]. The homology modeling was done based on the sequence alignment (Figure S1 and Figure S2), using different template structures using the design option in a new version of FoldX 2.8 [30],[31]. During this design procedure, FoldX is testing different rotamers and allows neighbor side chains to move. After reconstruction, all models have been passed through an additional optimization step by using the repair function of FoldX (detailed description in [33],[34]). Energy calculations of Ras-effector complexes have been done using FoldX as described before (http://fold-x.crg.es) [30],[31].
Simulations of a Minimal Ras-Effector Network
A model was generated based on previous models of EGF signal transduction (see Table S6). Simulations were performed using the SmartCell software (http://www.smartcell-crg.es) [60] using ordinary differential equations.
Supporting Information
Figure S1.
Alignment of UB domains
Figure S2.
Alignment of Ras proteins
Figure S3.
Diagrams of DGint and DGkon FoldX values plotted against the divergence time
Figure S4.
FoldX mutational scanning of RafRBD and RalGDS-RA. (A) Ras-RafRBD (pdb-entry 1GUA). (B) Ras-RalGDS (pdb entry 1LFD). Effect of all residues in Raf-RBD or RalGDS-RA on the contribution of DG kon as calculated by FoldX. Either positively charged residues were mutated to alanine (red) or negatively charged or neutral residues were mutated to lysine using FoldX and the difference compared to the WT DGkon was calculated and plotted for every amino acid position.
Table S1.
Experimental and calculated association rate constants
Table S2.
Divergence times and sequence identities for Ub domains
Table S3.
Template structures used for homology modelling
Table S4.
FoldX results for all homology models
Table S5.
Mean values and STDEV for homology models for ortholog complexes
Table S6.
Modelling parameters
Author Contributions
Conceived and designed the experiments: LS. Performed the experiments: CK DA. Analyzed the data: CK DA LS. Wrote the paper: CK LS.
- 1. Block C, Janknecht R, Herrmann C, Nassar N, Wittinghofer A (1996) Quantitative structure-activity analysis correlating Ras/Raf interaction in vitro to Raf activation in vivo. Nat Struct Biol 3: 255–251.
- 2. Pearce KHJ, Cunningham BC, Fuh G, Teeri T, Wells JA (1999) Growth hormone binding affinity for its receptor surpasses the requirements for cellular activity. Biochemistry 38: 81–89.
- 3. Piehler J, Roisman LC, Schreiber G (2000) New structural and functional aspects of the IFN-receptor interaction revealed by comprehensive mutational analysis of the binding interface. J Biol Chem 275: 40425–40433.
- 4. Wang Y, Shen BJ, Sebald W (1997) A mixed-charge pair in human interleukin 4 dominates high-affinity interaction with the receptor α chain. Proc Natl Acad Sci U S A 94: 1657–1662.
- 5. Batista FD, Neuberger MS (1998) Affinity dependence of the B cell response to antigen: a threshold, a ceiling, and the importance of off-rate. Immunity 8: 751–759.
- 6. Foote J, Eisen HN (1995) Kinetic and affinity limits on antibodies produced during immune response. Proc Natl Acad Sci U S A 92: 1254–1256.
- 7. Botti SA, Felder CE, Sussmann JL, Silman I (1998) Electrotactins: a class of adhesion proteins with conserved electrostatic and structural motifs. Protein Eng 1: 415–420.
- 8. Vetter IR, Wittinghofer A (2001) The guanine nucleotide-binding switch in three dimensions. Science 294: 1299–1304.
- 9. Bourne HR, Sanders DA, McCormick F (1990) The GTPase superfamily: a conserved switch for diverse cell functions. Nature 348: 125–132.
- 10. Bourne HR, Sanders DA, McCormick F (1991) The GTPase superfamily: conserved structure and molecular mechanism. Nature 349: 117–127.
- 11. Rapp UR, Goldsborough MD, Mark GE, Bonner TI, Groffen J, et al. (1983) Structure and biological activity of v-raf, a unique oncogene transduced by a retrovirus. Proc Nat Acad Sci U S A 80: 4218–4222.
- 12. Vojtek AB, Hollenberg SM, Cooper JA (1993) Mammalian Ras interacts directly with the serine/threonine kinase Raf. Cell 74: 205–214.
- 13. Rodriguez-Viciana P, Warne PH, Dhand R, Van Haesebroeck B, Gout I, et al. (1994) Phosphatidylinositol-3-OH kinase as a direct target of Ras. Nature 370: 527–532.
- 14. Hofer F, Fields S, Schneider C, Martin GS (1994) Activated Ras interacts with the Ral guanine nucleotide dissociation stimulator. Proc Natl Acad Sci U S A 91: 11089–11093.
- 15. Kikuchi A, Demo SD, Ye Z-H, Chen Y-W, Williams LT (1994) RalGDS family members interact with the effector loop of ras p21. Mol Cell Biol 14: 7483–7491.
- 16. Kuriyama M, Harada N, Kuroda S, Yamamoto T, Nakafuku M, et al. (1996) Identification of AF-6 and Canoe as putative targets for Ras. J Biol Chem 271: 607–610.
- 17. Emerson SD, Madison VS, Palermo RE, Waugh DS, Scheffler JE, et al. (1995) Solution structure of the Ras-binding domain of c-Raf-1 and identification of its Ras interaction surface. Biochemistry 34: 6911–6918.
- 18. Huang L, Weng X, Hofer F, Martin GS, Kim SH (1997) Three-dimensional structure of the Ras-interacting domain of RalGDS. Nat Struct Biol 4: 609–615.
- 19. Geyer M, Herrmann C, Wohlgemuth S, Wittinghofer A, Kalbitzer HR (1997) Structure of the Ras-binding domain of RalGEF and implications for Ras binding and signaling. Nat Struct Biol 4: 694–699.
- 20. Walker EH, Perisic O, Ried C, Stephens L, Williams RL (1999) Structural insights into phosphoinoside 3-kinase catalysis and signaling. Nature 402: 313–320.
- 21. Steiner G, Kremer W, Linnemann T, Herrmann C, Geyer M, et al. (2000) Sequence-specific resonance assignment of the Ras-binding domain of AF6. J Biomol NMR 18: 73–74.
- 22. Gronwald W, Huber F, Grünewald P, Spörner M, Wohlgemuth S, et al. (2001) Solution Structure of the Ras-binding domain of the protein kinase Byr2 from Schizosaccharomyces pombe. Structure 9: 1029–1041.
- 23. Nassar N, Horn G, Herrmann C, Scherer A, McCormick F, et al. (1995) The 2.2 Å crystal structure of the Ras-binding domain of the serine/threonine kinase c-Raf1 in complex with Rap1A and a GTP analogue. Nature 375: 554–560.
- 24. Nassar N, Horn G, Herrmann C, Block C, Janknecht R, et al. (1996) Ras/Rap effector specificity determined by charge reversal. Nat Struct Biol 3: 723–729.
- 25. Huang L, Hofer F, Martin GS, Kim SH (1998) Structural basis for the interaction of Ras with RalGDS. Nat Struct Biol 5: 422–426.
- 26. Vetter IR, Linnemann T, Wohlgemuth S, Geyer M, Kalbitzer HR, et al. (1999) Structural and biochemical analysis of Ras-effector signaling via RalGDS. FEBS Lett 451: 175–180.
- 27. Pacold ME, Suire S, Perisic O, Lara-Gonzalez S, Davis CT, et al. (2000) Crystal structure and functional analysis of Ras binding to its effector phosphoinoside 3-kinase gamma. Cell 103: 931–943.
- 28. Scheffzeck K, Grünewald P, Wohlgemuth S, Kabsch W, Tu H, et al. (2001) The Ras-Byr2RBD complex: structural basis for Ras effector recognition in yeast. Structure 9: 1043–1050.
- 29. Bunney TD, Harris R, Gandarillas NL, Josephs MB, Roe SM, et al. (2006) Structural and mechanistic insights into ras association domains of phospholipase C epsilon. Mol Cell 21: 495–507.
- 30. Guerois R, Nielsen JE, Serrano L (2002) Predicting changes in the stability of proteins and protein complexes: a study of more than 1000 mutations. J Mol Biol 320: 369–387.
- 31. Schymkowitz J, Borg J, Stricher F, Nys R, Rousseau F, et al. (2005) The FoldX web server: an online force field. Nucleic Acids Res 33: W382–W388.
- 32. Selzer T, Albeck S, Schreiber G (2000) Rational design of faster associating and tighter binding protein complexes. Nat Struct Biol 7: 537–541.
- 33. Kiel C, Wohlgemuth S, Rousseau F, Schymkowitz J, Ferkinghoff-Borg J, et al. (2005) Recognizing and defining true Ras binding domains II: in silico prediction based on homology modelling and energy calculations. J Mol Biol 348: 759–775.
- 34. Kiel C, Foglierini M, Kuemmerer N, Beltrao N, Serrano L (2007) A genome-wide Ras-effector interaction network. J Mol Biol 370: 1020–1032.
- 35. Kiel C, Serrano L (2006) The ubiquitin domain superfold: structure-based sequence alignments and characterization of binding epitopes. J Mol Biol 355: 821–844.
- 36. Orengo CA, Jones DT, Thornton JM (1994) Protein superfamilies and domain superfolds. Nature 372: 631–634.
- 37. Berg OG, von Hippel PH (1985) Diffusion-controlled macromolecular interactions. Annu Rev Biophys Biophys Chem 14: 131–160.
- 38. Northrup SH, Erickson HP (1992) Kinetics of protein-protein association explained by Brownian dynamic computer simulation. Proc Natl Acad Sci U S A 89: 3338–3342.
- 39. Vijayakumar M, Wong K-Y, Schreiber G, Fersht AR, Szabo A, et al. (1998) Electrostatic enhancement of diffusion-controlled protein-protein association: comparison of theory and experiment on barnase and barstar. J Mol Biol 278: 1015–1024.
- 40. Camacho CJ, Weng Z, Vajda S, DeLisi C (1999) Free energy landscapes of encounter complexes in protein-protein association. Biophys J 76: 1166–1178.
- 41. Camacho CJ, Kimura SR, DeLisi C, Vajda S (2000) Kinetics of desolvation-mediated protein-protein binding. Biophys J 78: 1094–1105.
- 42. Selzer T, Schreiber G (2001) New insights into the mechanism of protein-protein association. Proteins 45: 190–198.
- 43. Kiel C, Selzer T, Shaul Y, Schreiber G, Herrmann C (2004) Electrostatically optimized Ras-binding Ral guanine dissociation stimulator mutants increase the rate of association by stabilizing the encounter complex. Proc Natl Acad Sci U S A 101: 9223–9228.
- 44. Kiel C, Serrano L (2007) Affinity can have many faces: Thermodynamic and kinetic properties of Ras-Effector Complex formation. Curr Chem Biol 1: 215–225.
- 45. Sydor JR, Engelhard M, Wittinghofer A, Goody R, Herrmann C (1998) Transient kinetic studies on the interaction of Ras and the Ras-binding domain of c-Raf-1 reveal rapid equilibrium of the complex. Biochemistry 37: 14292–14299.
- 46. Linnemann T, Geyer M, Jaitner BK, Block C, Kalbitzer HR, et al. (1999) Thermodynamic and kinetic characterization of the interaction between the Ras binding domain of AF6 and members of the Ras subfamily. J Biol Chem 274: 13556–13562.
- 47. Linnemann T, Kiel C, Herter P, Herrmann C (2002) The activation of RalGDS can be achieved independently of its Ras binding domain: implications for an activation mechanism in Ras effector specificity and signal distribution. J Biol Chem 10: 7831–7837.
- 48.
Kiel C (2003) Investigation of Ras-effector complexes with changed electrostatic properties. Ruhr-Universitaet Bochum, Germany. Ph.D. thesis.
- 49. Hubbard TJ, Aken BL, Beal K, Ballester B, Caccamo M, et al. (2007) Ensembl 2007. Nucleic Acids Res 35: D610–D617.
- 50. Remm M, Storm CEV, Sonnhammer ELL (2001) Automatic clustering of orthologs and in-paralogs from pairwise species comparisons. J Mol Biol 314: 1041–1052.
- 51. O'Brien KP, Remm M, Sonnhammer ELL (2005) Inparanoid: a comprehensive database of eukaryotic orthologs. Nucleic Acids Res 33: D476–D480.
- 52. Schultz J, Milpetz F, Bork P, Ponting CP (1998) Smart, a simple modular architecture research tool – identification of signalling domains. Proc Natl Acad Sci U S A 95: 5857–5864.
- 53. Letunic I, Goodstadt L, Dickens NJ, Doerks T, Schultz J, et al. (2002) Recent improvements to the SMART domain-based sequence annotation resource. Nucleic Acid Res 30: 242–244.
- 54. Kholodenko BN, Demin OV, Moehren G, Hoek JB (1999) Quantification of short term signaling by the epidermal growth factor receptor. J Biol Chem 274: 30169–30181.
- 55. Schoeberl B, Eichler-Jonsson C, Gilles ED, Mueller G (2002) Computational modeling of the dynamics of the MAP kinase cascade activated by surface and internalized EGF receptors. Nat Biotechnol 20: 370–375.
- 56. Yamada S, Taketomi T, Yoshimura A (2004) Model analysis of difference between EGF pathway and FGF pathway. Biochem Biophys Res Commun 314: 1113–1120.
- 57. Sasagawa S, Ozaki Y, Fujita K, Kuroda S (2005) Prediction and validation of the distinct dynamics of transient and sustained ERK activation. Nat Cell Biol 7: 365–373.
- 58. Kiyatkin A, Aksamitiene E, Markevich NI, Borisov N, Hoek JB, et al. (2006) Scaffolding protein Grb2-associated binder 1 sustains epidermal growth factor-induced mitogenic and survival signaling by multiple positive feedback loops. J Biol Chem 281: 19925–19938.
- 59. Birtwistle MR, Hatakeyama M, Yumoto N, Ogunnaike BA, Hoek JB, et al. (2007) Ligand-dependent responses of the ErbB signaling network: experimental and modeling analyses. Mol Syst Biol 3: 144.
- 60. Ander M, Beltrao P, Di Ventura B, Ferkinghoff-Borg J, Foglierini M, et al. (2004) SmartCell, a framework to simulate cellular processes that combines stochastic approximation with diffusion and localisation: analysis of simple networks. Syst Biol 1: 129–138.
- 61. Wang DYC, Kumar S, Hedges SB (1999) Divergence time estimates for the early history of animal phyla and the origin of plants, animals and fungi. Proc Biol Soc 266: 163–171.
- 62. Hedges SB (2002) The origin and evolution of model organisms. Nat Rev Genet 3: 838–849.


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It's a meditation, as affecting as it is entertaining, on the limits of violence and the power of unchained empathy. show more
For those who gripe that America doesn't make cars or movies like it used to, Clint Eastwood has two words for you: Gran Torino. show more
A lifetime in movies runs through this prime vintage Eastwood performance. You can't take your eyes off him. The no-frills, no-bull Gran Torino made my day. show more

subset: commoncrawl


text:
Four police officers were injured and several dozen people were arrested in protests in which shop windows, including that of McDonald's on Wenceslas Square, were smashed.
The demonstrations, which also targeted a Kentucky Fried Chicken shop, were against the growing influence of multinational corporations and their effect on the environment. They began with an open air rock concert billed as the "Global Street Party '98", timed to coincide with the G8 summit in Birmingham, and ended with a punch-up in Wenceslas Square. This square was the cradle of the 1989 "velvet revolution" where a quarter of a million Czechs gathered and in the course of a week peacefully brought down their communist government.
In the years after 1989, Prague underwent a renaissance - politically, under its world-famous playwright president, Vaclav Havel, and economically. It became a magnet, too, for Western youth. What Paris had been to bohemian Americans in the 1930s, Prague became in the 1990s.
The once-quiet Charles Bridge, one of the city's most famous and elegant landmarks, became so crowded at all hours of the day with thousands of tourists and temporary residents, that it was scarcely possible to glimpse the bridge itself.
Czechs were not always enthusiastic about all the changes that were taking place. The tourism boom brought millions of pounds of much needed foreign currency, as Prague became a number-one destination in the holiday brochures. But Czechs themselves often felt left out of the loop. Radical economic change brought high unemployment and poverty. There was the constant tension, too, between the need to do everything possible to bring foreign money into the city and the need to preserve the distinctive character of Prague.
And while communist repression has gone, there have been big cuts in education and health spending, and there are homeless people on the streets.
The weekend violence came as several thousand people, protesting over these changes as much as alleged damage to the environment perpetrated by multinationals, left the concert and marched on the city centre. A small group hurled paving stones and broke the glass windows of McDonald's - the third such attack.
Mainstream environmental groups were quick to condemn the violence. Greenpeace regretted that the protest had been described as ecological, "because that damages the image of preserving nature and the environment". The chairman of the environmental pressure group Duha complained: "This is abusing the name of the environmental movement."
Martin Bursik, the Environment Minister, also drew a line between the demonstrations and other environmental groups. He invited Green organisations for a meeting yesterday, and argued: "Nobody can seriously think that the programme of the ecological movement equals looting and stealing salami."
- More about:
- Czech Republic
- Mcdonald's
- Prague


text:
Government tenders for £1bn hosting framework
The Government Procurement Service has issued a tender for a hosting services framework worth between £100m and £1bn
This Article Covers
Government & public sector IT
The Government Procurement Service has put out a framework tender for hosting services worth between £100m and £1bn.
The four-year framework for IT services will include consulting, software development, internet and support. It will have a minimum of 12 participants and it will be used across central and local government, said the Government Procurement Service.
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The public sector is in need of hosting services for large and complex application environments, it said.
"This procurement is targeted at provision of the applications and services required to make available the government's large, mission critical high availability services that utilise complex infrastructure with server or mainframe based technology typically hosted in one or more traditional data centre environments," said the Government Procurement Service.
The framework is designed to drive towards standardisation in accordance with the published Government ICT strategy, said the tender.
No further details were provided about the specific services the Government Procurement Service is seeking. The tender deadline is 20 August 2012.
The note follows a recent £4bn framework award for IT hardware across the public sector, which includes desktops, laptops, tablets, servers, printers and most commonly used peripherals.
Enjoy the benefits of CW+ membership, learn more and join.
European law enforcement seeking smart ways to fight cyber crime
Read more on IT for government and public sector
Gov.uk Verify ID providers eye the prize of HMRC's digital users – but what happens without them?
HMRC ID vs Gov.uk Verify – what's the difference, and why it matters
GDS, HMRC and Verify: so much for cross-government digital collaboration
The government's transformation strategy is ambitious, but is it achievable?


text:
Wolf game
By SEJER
Open iTunes to buy and download apps.
Very popular among the French audience, "The Wolf game" is finally available on iPhone.
Discover or rediscover a best-seller game for the 3/6 year olds.
"Quick! Fill your basket before the Wolf is completely dressed! Click on the magic token and a strawberry, a flower or another object will pop up. But if it's the Wolf's head, beware! He puts his hat on!"
Beautiful pictures, funny animations and amusing voice-overs in a playful and non-competitive game: 2 playing modes (team versus Wolf ; Who will Win?), basic rules, short rounds to be played by one or in team players.
The original board game has been aprouved by a large amount of parents and families.
"A very simple game, very easy to use, we can play together against the wolf, a nice way to softly learn how not to be always the winner"
"A lot of fun for little daughter, very short rounds, very easy to understand!
All the family loves it!"
What's New in Version 1.1
The application "Jeu du loup" will now work with former iOS devices.
iPhone Screenshots
- Category: Games
- Updated: 30 March 2011
- Version: 1.1
- Size: 13.6 MB
- Languages: English, French
- Developer: Sejer
- © Nathan, 2014
Compatibility: Requires iOS 3.0 or later. Compatible with iPhone, iPad, and iPod touch.


text:
We're sorry, this listing is off the market. Try these listings instead:
Consulting Service #224203 posted Jan 26, 2011
- Bangkok, Bangkok, Thailand
- $75,000
Established five years ago, this Bangkok based Consultancy firm has over 70 clients on the books, including some of Thailand's leading corporations, car manufacturers, universities, banks, technological institutes and website developers. Providing a wide range of consultancy services, the company helps...
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- Doha, Baladīyat ad Dawḩah, Qatar
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Grade A architect and engineering firm operating in the market for more than 20 years whereby designed and supervised construction of millions of square meters through literally hundreds of projects that has been constructed with total cost exceeds QAR 15 billion and known as a major landmark projects...
Training in Live Sound Engineering #394191 posted Nov 13, 2015
- London, England, United Kingdom
- Available On Request
Company Address-Scala 27 Kings Cross, London England. Postal Address-N19NL. FINANCIAL DETAILS: Asking Price-Undeclared Turnover-£110,000 Gross profit-£40,000 Net profit-£20,000 Premises-Freehold
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text:
Barack Obama's low-key endorsement of the BlackBerry during his presidential campaign was estimated to be worth as much as $50m (£31m). But as the aspirational status of the BlackBerry has slipped, despite recent models incorporating the swipe-screen technology that helped make Apple's iPhone so attractive, a new crowd has emerged to adopt the beleaguered BlackBerry. Not so much early-adopters as latecomers, they may well be the future for one of the mobile world's most famous brands.
It happened almost by accident at first, then decidedly on purpose. The device is now popular among America's youth, and the same is happening in Britain, and the key is free messaging.
BlackBerry-maker Research in Motion saw the evaporation of corporate interest coming for a while and has battled the almost inevitable downturn with aggressive marketing to teenagers. Trend watchers noted the BlackBerry's popularity among kids in Hollywood a couple of years ago and predicted its spread; anyone sitting on a bus at 4pm on a weekday will see that they've been proved right. Unlike the iPhone, it's possible to pick up a BlackBerry for free on a sub-£20 price plan, and while this may not make it a prestige device, kids don't care. It's cheap. And, more importantly, it has BlackBerry's proprietary messaging service, BBM.
BBM only works between BlackBerry devices, is more sophisticated and flexible than the text message and is, crucially, free to use.
Parents of teenagers may have witnessed its power. The cool girl at school gets a BlackBerry Pearl, possibly the pink model. Her friends follow suit, and they communicate over BBM. Boys, not wanting to be left out of the conversation, get a BlackBerry, too, and so it spreads. This may not have been the firm's intention, but it has worked out brilliantly. The kids love it. The question is whether enough of them will embrace the BlackBerry to offset the deserting suited clientele.Reuse content


text:
Yes, there are differences in Verbenas, but there are other lines that
would perform as well as tukanas. I think PW has more than one line of
Was there something in Jim's photos re the verbenas?
-------Original Message-------
From: email@example.com
Sent: 05/29/03 06:08 PM
To: email@example.com
Subject: Re: [CHAT] Plant Labels
> In a message dated 5/29/03 9:33:18 AM, firstname.lastname@example.com writes:
<< This one garden specified it had to be tukana
hybrid verbenas, not just any verbenas. Finally found one place that had
them, had to pay $5 each. Could have gotten much nicer plants in better
condition, similar color for $3 each. >>
Kitty, is that a proven winner? If so, it probably had the big labels in
addition to the big price. I must agree, however, there is a definite
in the performances of verbena.
If you have weeds, you don't have enough plants.
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text:
The Crossroads of the Aether
207 members132 Comments 10 Likes
139 members52 Comments 21 Likes
195 members142 Comments 29 Likes
Mlle. K. Harte, Danser du Ventre has not received any gifts yet
Welcome to the Empire Ms. Hart! Did you enjoy Comicpalooza? If you wish you can follow this link to the 42nd Airship Battalion-- join if you like. Enjoy the Empire.
© 2017 Created by Hephzibah Marsh. Powered by


text:
Dodgers hire McGwire as hitting coach
The Dodgers, as expected, hired Mark McGwire as their hitting coach today.
McGwire, who spent the past three seasons as the St. Louis Cardinals' hitting coach, apparently wanted to work closer to his home in Irvine.
"Mark has been able to translate his success as a big league hitter into his work as hitting coach with the Cardinals," Dodgers general manager Ned Colletti said in a statement. "He earned the respect of both veterans and young players while in St. Louis, and the offensive numbers the Cardinals produced under his tutelage speak for themselves.
"We're excited to have him join our coaching staff and anticipate great things from his work with our hitters in 2013 and beyond."
McGwire was celebrated during his career for his power hitting and finished with 583 home runs. But he also drew severe criticism when he refused to answer questions about steroid use during his playing career when called to appear at a congressional hearing in 2005.
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text:
Novum Hotel Silence Garden Cologne Brueck
No prices found through our partners. Please contact the business directly or check some of our recommended alternatives.
How has this hotel rated in the past?
- Drycleaning onsite
- Breakfast Available
- Wireless internet connection in room (free)
- Non-smoking hotel
- High-speed internet in room
- Non-smoking rooms
Photos of Novum Hotel Silence Garden Cologne Brueck
- Breakfast Available
- Wireless internet connection in room (free)
- Non-smoking hotel
- High-speed internet in room
- Non-smoking rooms
- Free High-Speed Internet
- Free parking
- Drycleaning onsite
- Meal plan
TripAdvisor Reviews Novum Hotel Silence Garden Cologne Brueck
Travel Blogs from Cologne
... We stayed clear of the left lane for the most part and did remark that three hot machines blew by us(on each others tail) as if we were standing still and we were doing 130 kmh. Some things never change!!!
As I write, we are enjoying a refreshment in our Hotel zum Muehle located on the Rhine River. A wonderful setting but not enough good light for pics when we arrived. Tomorrow after breakfast (our standard routine) will work much better. We have ...
Wonderful location
... for a table and having two beers and leaving for the next pub. Home at 10:30 or the ship leaves!! We had our friends Carol and Rob in our group so that made it fun. It was so loud everywhere we couldn't hear the guide. All the locals were so young and we looked like a group of fish out of water...or beer, as it were...We should have gone back to Papa Joe's and just listened to the fake musicians and soccer fans! ...
Early this morning, about 4.30, we sailed passed Frankfurt and onto Cologne, berthing at about 1pm. We opted for the city walking tour which was about 1.5 hour duration. We learnt about the history of Cologne, or Koln as it is called in Germany. With a population of 1 million, it's the 4th biggest city in Germany (behind Berlin, Hamberg, Munich). The Romans established a settlement at this location on the Rhine about 2000 years ago and it has has continuous settlement ever ...
Day 13 Cologne - Germany.
After spending the evening in Bonn, which gave the crew the opportunity of a welcome night out, if they chose, we set sail for Cologne at 5.15 this morning and docked at 8am. Obviously it was just a short trip and I don't know the reason why we didn't continue here last night. Cologne is a large city, and to me in the category of Vienna, Nurenburg and Budapest. They are no doubt nice for various reasons, but to me do not have the character and appeal of ...
My Last Days in Germany
... about a series of subjects―his family (wife had passed away last year), professions, my time in Germany, his relatives in the U.S. (Toledo and Philadelphia) and a bit of politics (U.S. and German). Afterwards I retrieved my luggage and took it to the lobby and paid my bill. I made the five
minute walk to the bus stop, purchased my ticket and awaited the #201 bus to the train station, where I arrived about 30 minutes prior ...


text:
John's wine culture experience began with Brand's "Laira" Coonawarra reds at the age of 20 years in 1973.
Thereafter, the 'wine bug' had struck, and the symptoms manifested as a collection of Australian and European wines.
In 1993 we planted three acres each of Chardonnay and Pinot Meunier in our desertscape backyard at Greenhills, Victor Harbor, Southern Fleurieu Peninsula. Our first sparkling wine was released late December 2002.
1998 saw John and a good mate 'pick over' a machine harvested vineyard at Lyndoch. Just under a tonne of mixed red grapes produced a 'mates quaffer'.
In 1999 John accepted an offer of a tonne each of 80 year old dry grown Grenache and 5 year old Shiraz from the same vigneron and David Powell of Torbreck offered to vinify the small parcel of fruit.
Mt Billy was born.
The Mt Billy Wines website makes use of Macromedia Flash software. To experience it the way we intended, you need Macromedia Flash Player, a plug-in for your Web Browser that can play the content we've created.
Why not download and install it now? It's easy, and you'll be joining more than 700 million people who can experience musical tracks, sound effects and animations integrated with innovative interfaces.
If you are sure you have the correct version of flash installed you can view the site by clicking here


text:
Office 365 and Microsoft office 2013 could be the newest office tools from Microsoft. There could be several unexpected concern wherever messages or uninstall process of Office 2013 ceases sudden before achievement, and then caught forever can take place. Inside the guidebook, the Office ideas are organized into classes, for example Software Basics" and Report Management." Each set of tips about a topic, including Using the Ribbon" or Protecting Papers," is designated and organized for clarity and order of route. Below is a good example of the tips under Using the Ribbon" in the Interface Basics" group. While both bedrooms offer you a diverse pair of purposes and alternatives, the variation of Perspective 2013 contained in either suite is exactly the same.
All other star signs for example From Workplace communications, delivery statements and reacted/forwarded but additionally calendar meeting announcements remain being shown on the right above time or information. To supply a far more streamlined knowledge without the need of popping up added concept windows and dialogs anymore, Prospect 2013 features in-line answer that allows you to push response and right start publishing your answer within the Reading Pane (which means that it is not only a Reading Pane anymore and they must have modified the brand).
If you've taken the dive together with the fresh-look hint- programs that are friendly, these recommendations and tips can help you get further using the application - from creating the interface to embedding films, there is lots to discover. We shall give attention to the brand new features in Office 2013 (although you'll locate some methods that work over the panel), while delivering a mix of quick suggestions and more in-depth ideas to accommodate every degree of user. Office Next - the Cloud along with Office - Learn how a new Workplace is to the cloud to produce production possible across numerous gadgets.
By demonstrating just one point under the headline Prospect 2013's default means of presenting a note is. View will come in new colors now - instead of the violet, dark and silver subjects that you may 've got used to, you can now modify the topic to Light Dull or Dark Gray, for retaining it Bright when you also can opt. You pick the arrow then click down close to Workplace Theme as a way to select one of the three possibilities and can go-to Report to take action. As a way to achieve this in Outlook 2013, your Diary, press right merely opens and goto shade. microsoft office 2013 activator
Last updated 658 days ago by johnson lovell


text:
I've got a few varieties growing here in my sandy soil. Never had a problem
with them until this winter which killed off two old bushes. I'm replacing
them with newer cultivars.
They do tend to leave offspring behind. I should qualify that statement,
'White Profusion' has been seeding around. The others are not as
Long Island, NY
Zone 7a (Average min temp 50 - 00)
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text:
Comfort Suites Gwinnett Place
No prices found through our partners. Please contact the business directly or check some of our recommended alternatives.
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Travelers also recommend:
- More recommendations
TripAdvisor Reviews Comfort Suites Gwinnett Place Duluth
Travel Blogs from Duluth
Last Day in Atlanta
Today is the last day of the Conference and our last full day in Atlanta. Drive to Orlando tomorrow to join the cruise on the 11th. I left the hotel early and caught the shuttle bus to the Martin Luther King Jnr Historic site. It included the Ebeneza Baptist Church that his father and he were ministers. The church was the centre of his civil rights work. His wife, after his assassination, took over and continued his amazing work for peace ...
Unpacking equipment and yoga!
This morning the group had a CPR session. Since I'm already certified and need a health provider version instead of the commoner version that was offered (hehe), I spent the day unpacking the forensic science supplies and getting materials together. I got everything on my list and am thrilled!! They really hooked me up with everything on my supply list. Then in ...
Holiday with family
... enjoyed spending time with Ellie and Lola round the pool, though Lola kept her distance near the water!,
Our trip to the zoo took us on the local train system, most efficient. Then a wee stroll up to the zoo, this day was extremely hot so the walking was a little ******* Ellie, all of us if we were honest. A well kept zoo, and plenty to see though we did not get to see the giraffes as their enclosure was being revamped. But we did get to see a set ...
Just Passin' Through (At least we hope..)
... behind the gate agent, and it was lit up all red! I couldn't hear what was happening but I saw another man approach, saw Jack taking his **********, the pat down, more rubbing of stridex pads, his bags separated. And there at his feet, sat the culprit of the search. My backpack! What?
He's talking to them, they process more pads, more red. He looks confused. And it hits me! Just three weeks before this, I carried that backpack to ...
Baltic Cruise
... industrial. Gdansk shipyard is no longer in use but is of historical significance because of the labor uprising in 1980 lead by Lech Wallensa. Our tour guide Elizabeth was extraordinary. She was nonstop in her explanations of the sites we were passing and visiting. We passed through Gdynia, Sobort(the resort city), and stopped in the old city of Gdansk. Gdansk was almost totally decimated in WWII. This was the siege that began WWII. After the war the Russians rebuilt the city to ...


text:
1. Kimberly Gordon is known as "flash Gordon" to her parents because she was born just after her mother was struck by lightning.
2. The family claims their child still has permanently static hair a year after the incident.
3. The young girl was delivered by emergency caesarean section after Kendra Villanueva was hit during a rainstorm in Albuquerque, New Mexico.
The pair, and Kimberly's father, Ian Gordon, were rushed to hospital after being struck.
4. Kimberly's parents say she still can't digest food properly or sit up unaided.
Villanueva told KOAT: 'It's hard seeing that she can't eat like she's supposed to and can't have food yet, just through the [feeding] tube. But other than that I'm glad she lived through it all."
"She's just kind of a play-it-by-ear baby," Gordon added.
"Everybody we've seen says it's a miracle she's alive."


text:
Appetizer Meat Balls
28 May 2009
I was on a hunt for appetizer recipes and bumped into this Appetizer Meat Ball. Tried this for the party last week and believe me when I say that it was a hit! Why don't you try these easy to make Appetizer Meat Balls and revel in the accolades that flow in.
|Ground beef
|3⁄4 Pound
|Ground pork
|1⁄4 Pound
|Rolled oats
|3⁄4 Cup (12 tbs)
|Chopped water chestnuts
|1⁄4 Cup (4 tbs)
|Worcestershire sauce
|1⁄4 Teaspoon
|1⁄2 Cup (8 tbs)
|Onion salt
|1⁄2 Teaspoon
|Garlic salt
|1⁄2 Teaspoon
|Tabasco sauce
|3 Drop
|1 Cup (16 tbs) (For Browning)
Blend all ingredients; form into about 75 small balls.
Brown well in margarine; drain on paper towels.
Rate It
Nutrition Rank
Nutrition Facts
Serving size: Complete recipe
Calories 3367 Calories from Fat 2673
% Daily Value*
Total Fat 299 g460.6%
Saturated Fat 76 g380%
Trans Fat 0 g
Cholesterol 332.9 mg111%
Sodium 1926.9 mg80.3%
Total Carbohydrates 81 g27%
Dietary Fiber 11.3 g45.2%
Sugars 8.3 g
Protein 96 g191.9%
Vitamin A 157.8% Vitamin C 2.4%
Calcium 19.5% Iron 48.9%
*Based on a 2000 Calorie diet
Appetizer Meat Balls Recipe


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THE WALL is written and directed by the team that brought you Zombie Dearest, one of the smartest low-budget horror/comedies of the last five years… Two-time Best Feature Winner, Two-time Audience Favorite winner, Zombie Dearest was embraced by festivals, critics and filmmakers alike and is featured as one of the very few recommended picks in the soon-to-be released "Zombie Movie Encyclopedia 2000-2010" by Peter Dendle.
Still, you might never have heard of Zombie Dearest. That's because with little independent filmmakers like us, every penny goes directly into the production. We won't be saturating the airwaves with ads. Still, we think THE WALL is the real deal, a shot at success the old-fashioned way… a great film, through festivals and word of mouth, finding its audience.


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Lot 35: Taino Recumbent Skeletal Frog Man (1000-1500 CE)
October Estates
by Material Culture
October 31, 2016
Philadelphia, PA, USALive Auction
Looking for the realized and estimated price?
Description: Taino Recumbent Skeletal Frog Man (1000-1500 CE). The recurved base suggests it may have been worn on arm. Eyes are raised in slits denoting a dream state or trance. Made of basalt. 4"L x 1.75"W, (10 x 4 cm).
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Magnaflow Performance Exhaust 17938 Pro Series Diesel Performance Exhaust
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Summary | Excerpt | Reviews | Beyond the book | Readalikes | Genres & Themes | Author Bio
The publisher's blurb about Trieste, the first novel published in English by Croatian author Daa Drndć, does not begin to convey the harrowing intensity of this riveting, singular work. The book illuminates the history of the Italian Jews of northern Italy under German SS occupation during World War II. This is filtered through the memories and experiences of Haya Tedeschi, an eighty-two-year-old Catholicized Jewish woman who was twenty when her infant son was taken away as part of Heinrich Himmler's Lebensborn program.
It is now 2006 and at long last, Haya is looking forward to meeting her son in her hometown of Gorizia, Italy, a town that borders Slovenia. Gorizia, a few miles north of Trieste, is where Haya has lived since the war. Throughout the course of the sixty-two years since five-month-old Antonio Tedeschi, bastard son of a high-ranking SS officer, was ...
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Separate names with a comma.
Discussion in 'The Anime Forum' started by Takao, Nov 6, 2012.
Nicktoons hasnt seen the last of DBZ Just yet!
Interesting. Must have done better in America then they expected.
Now my one reason for disliking Kai is gone. With the Buu arc, Kai watchers can finally see the full story.
I wonder if it was Saban Brands who put the money for DBZKai to continue. Funimation said it didnt sell very well for them.
Ugh. Argh. Ack. Well, at least the cast is recording it, but still...what a waste of money.
Its because Toei Animation does not have any other franchises to exploit internationally. One Piece, Pretty Cure,Zatch Bell all failed. Its actually an investment (Paying money to make money) when you think about it
Cool. At least it will be complete. Absolute waste of money, but it's not like Toei puts any effort into most actual new animation either. New Dragon Ball stuff does tend to look better, though. Seeing that TFS special today reminded me of that.
Well, now that I've had some breakfast it's time to sit back and see how the hell this all plays out. I wonder how they'll cut the ninety-one episodes down?
That had not escaped me.
If Saban indeed had a hand in funding this, they would've made sure it would broadcast exclusively on Vortexx. Can't have others profiting from your investment.
Considering Kai got impressive ratings on Nicktoons and now on Vortexx, I wouldn't call it a huge waste. Besides, it's nice that it won't stay incomplete.
Looks like Nagasaki Yukio might be returning to the series, too. Nagasaki was the Audio Director, in charge of placing music and directing the actors. Ugh.
Well, it was complete. Toei never said they were going to do the whole series.
So, I'm guessing this will be about 50 or so additional episodes?
So, Kai is getting the Buu saga? Great! When it comes on I'll be sure to let my little brother know so he can catch it.
We'll know whenever we know. The first arc was fifty-four episodes, the second was fourty-four. I'm hoping for fourty myself. If you're going to do this, do this was different from what Dragon Ball Z did as possible.
I'm not really surprised since not only was DBZ Kai such a huge hit for Nicktoons but it seems even DBGT is doing well for them. I'm guessing this is an idea they struck recently to get greenlit otherwise I'm sure they would of waited some BEFORE showing GT if they were going to get a Buu Kai. As for how they're going to do it and how many episodes it will be... hard to say. Maybe since this isn't a Japanese production like the rest of Kai they'll try catering more to western or other auidences and thus it could be more quicker paced or they could try stretching it out more then regular Kai since unless they want to just make totally original material there's not really much else they can do with DB besides Kai Buu.
I'm thinking this has a lot more to do with international markets but it would be interesting if Saban and/or Viacom helped foot the bill.
It's likely the international markets. Dragon Ball Kai has been licensed in a hell of a lot more countries than any other recent Japanese cartoon series and airs on the teley. Toei can guarantee strong licensing fees and royalties and won't have to do any marketing themselves, so they're a bit more free to produce another arc worth of episodes. Domestically, Kai is old hat, espicially with the Dragon Ball Z name now being brought back for the 2013 film.
Anyway, I did some number crunching. Tanaka said that they were recording the part where Goten and Trunks disguise themselves as Mighty Mask. I believe that happened in Dragon Ball Z episode #214. Assuming Kai adapts at least two episodes an episode, that'll be around episode #6 or #7.
1. episodes #200-201
2. episodes #202-205 (the way Dragon Ball Z adapted the comic here will require a bit of good editing to try to match it to the comic's story)
3. episodes #206-207.
4. episodes #208-209.
5. episodes #210-211.
6. episodes #212-213.
7. episodes #214-215.
Of course, I could be entirely wrong here, so who knows...
I remember a few comments that suggested that DBZ Kai could continue on outside of Japan after it ended, although I think most of them were about the possibility of Funimation editing the episodes together themselves. Still, it's surprising to see that this did happen. At least it will cover the rest of the series.
You're wrong about this part.


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Investors spent most of 2012 shedding stocks and ramping up on bonds, but many say this expansion into bonds, which has been going on for four years, is ready to end in 2013. The fear is that its bubble will burst. "Mathematically, it's next to impossible to get the kind of returns on bonds you've seen over the last few years," said Bank of America's Kate Moore.
Full Story:The New York Times (tiered subscription model)


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Tropical Fish Keeping - Aquarium fish care and resources (http://www.tropicalfishkeeping.com/forum.php)
- Beginner Freshwater Aquarium (http://www.tropicalfishkeeping.com/beginner-freshwater-aquarium/)
- - Are These Cardinal Tetra Eggs? (http://www.tropicalfishkeeping.com/beginner-freshwater-aquarium/these-cardinal-tetra-eggs-109615/)
Are These Cardinal Tetra Eggs?
1 Attachment(s)
Hey Guys,
Do these look like cardinal tetra eggs to you. Their all over the sides of the tank and I couldn't think of anything else.
I think the eggs are all those white things in front of the gourami.
Do you mean the cluster of opaque "eggs" directly in front of the gourami, at the very bottom of the photo (which i think is on its side)? Looks like a cluster attached to the tank wall. If these were fish eggs, the fish would likely be eating them fast. Cardinal tetra spawn in plants and scatter the eggs, so if these are eggs thy are almost certainly not from the tetra. Is that gourami eating them, or guarding them, or ignoring them?
Yes those are what I'm talking about. I don't have a lot of plants for cardinal tetras to spawn but still you're right they probably aren't cardinal eggs. The gourami is ignoring them. Probably just dust or dirt or something. Thanks for the post.
P.S., yeah the photos on its side.
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Game Winner Talisman Widget
by: luckyrune • 0
All Android applications categories
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by: luckyrune • 0
Increase your chances to win games by carrying this Talisman. This gambling spell allows the user to get better results when playing the lottery, card games, betting, and other games of chance. The first rune - Laguz - increases the gambler intuition, the second – Mannaz – gives experience, wisdom and skill in the game, while the third – Tiwaz – gives strength in battles.
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Professeurs de russe à Montreal >
Partagez ce profil :
Ville / Pays d'origine : Novossibirsk, Russia
Langue maternelle : Russe
Langues parlées : Anglais, Français, Russe
"Natalia Russian class is excellent. She has so much energy and enthusiasm, and is always very prepared. She is able to make my three year old daughter feel comfortable, and makes learning fun.""Kristelle, Montreal
"Listens closely and corrects mistakes. Provides interesting materials for reading practice and discussions. Lets me complete sentences before correcting. Good lessons."CB, Washington DC
"Well prepared and on time. Listens and corrects mistakes . Materials chosen are relevant and interesting."Christopher, Washington DC
"I really liked my experience with this language course, which helped me a lot with my reading, writing and speaking skills. Natalia gave me a lot of practical examples and situations, and managed to adapt her classes very well to work on what I had trouble with. This has been a really helpful course, and I will definitely continue learning Russian with Natalia in the future. "Rémy, Montreal
Hello! I am a certified teacher of Russian language holding professional Diploma of Pedagogical University (Russia). I possess 3 years of professional teaching experience from school, college and private practice.
I have a professional expertise in my mother language - Russian, and am happy to share its beauty and richness with my future students.
Bachelor`s Degree in Teaching Russian Language, Literature and English Language.
Barnaul Pedagogical University (Barnaul, Russia).
I taught Russian and English as a foreign language to adults and kids aged 3 to 15 in Novossibirsk (Russia) and Montreal (Canada).
Cours offerts par ce professeur :
Autres professeurs :


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All Entries Tagged With: "Citrix Systems"
Citrix Enables "Workshifting" For IT With New, Free GoToManage App for iPad
Posted by Mary Ann Neder | May 25, 2011 | Comments 0Today at Citrix Synergy, the event where virtual computing takes center stage, Citrix Systems announced GoToManage for iPad, a free application that enables users to securely troubleshoot and provide real-time technical support to a computer user's system from anywhere. Workshifting for IT will soon be possible with the availability of the number one remote support solution[1] on Apple's popular mobile media and computing device.
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Mortgage Rates Today and Refinance Rates: 30 Year Mortgage Rates and Refinance Rates Averaging 4.82%
Today's mortgage rates and refinancing rates are higher compared to yesterday's rates. 30 year mortgage rates and refinancing rates today are averaging 4.82 percent, up from yesterday's average 30 year mortgage rate and refinance rate of 4.69 percent. Conforming 30 year mortgage refinance rates and loan rates in New Jersey are slightly lower than the current national average rate. Current 30 year mortgage rates and refinance mortgage rates in New Jersey averaging 4.80 percent.
15 year mortgage rates and refinance mortgage rates today are currently averaging 4.14 percent, up from yesterday's average 15 year mortgage rate and refinance rate of 4.05 percent. 15 year refinance mortgage rates and mortgage interest rates in Ohio are slightly lower than the national average 15 year interest rate. Today's 15 year interest rates in Ohio are averaging 4.12 percent.
30 year jumbo mortgage rates and jumbo refinancing rates are averaging 5.38 percent, up yesterday's average 30 year jumbo rate and refi mortgage rate of 5.24 percent. 30 year jumbo mortgage loan rates and jumbo refinance loan rates in Michigan are lower than the national average rate. 30 year refinancing rates and jumbo mortgage rates in Michigan are averaging 5.31 percent.
15 year jumbo refinancing interest rates and jumbo rates today are averaging 4.49 percent, down from yesterday's average jumbo loan rate and 15 year jumbo refi rate of 4.69 percent. 15 year jumbo interest mortgage rates and refinance interest rates in Georgia are higher than the national average. 15 year mortgage rates and refinancing loan rates in Georgia are averaging 4.60 percent. Track mortgage rates by using our rates widgets.
Following is a more in-depth home mortgage loan rates report released by MonitorBankRates.com this past Monday:
Mortgage Rates Today & Refinance Rates Today
30 year conforming mortgage rates and refinance rates today are averaging 4.77 percent, up from last week's average 30 year home loan rates and refi rate of 4.74 percent.
Average Mortgage Rates
Recent Mortgage Rate Articles
Mortgage Rates Hit New Lows for 2014
Mortgage Rates Decline as Fed Continues to Taper
Mortgage Rates are Stable as Fed Drops Bond and MBS Purchases to $55 Billion a Month
Mortgage Rates: Average 30 Year Mortgage Rates Fall to 4.25 percent
Mortgage Rates Lower Due to Emerging Market Fears and Lower Bond Yields
Mortgage Rates Forcasted to Move Higher in 2014 and 2015
Mortgage Rates Move Lower Today: Best 15 Year Rates Just Under 3.00%
Current Mortgage Rates March Higher as Applications for Mortgages Decline
Today's Mortgage Rates Move Higher, 30 Year Refinance Rates at 4.38 Percent
Mortgage Rates: Lowest 30 Year Mortgage Rates Today at 3.625%
Mortgage Rates Decline as Home Affordability Slips
Mortgage Rates Today Lower After the Government Shutdown
Conforming 30 Year Mortgage Rates Heading Back Down to 4.00 Percent
Mortgage Rates Continue to Trend Lower as the Federal Government Shuts Down
Mortgage Rates at Credit Unions Decline on News the FOMC Won't Taper
Mortgage Rates Rise as Home Price Increases Slow
Current Higher Mortgage Rates Haven't Dampened Existing Home Sales
Mortgage Rates Take a Breather from Recent Gains, 30 Year Rates Averaging 4.31 Percent
Rising Mortgage Rates Will Cool the Housing Market
Current Bank Mortgage Rates Move Lower From Recent Highs, 30 Year Mortgage Rates at 4.35 Percent
Mortgage Calculator
Mortgage Rate Archives
Mortgage Rate
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More Spec, More Strat
A tonally versatile guitar with time-honored Fender style and sound, the Deluxe Strat has a few tricks up its sleeve. Special switching unlocks extra pickup combinations for those moments you find yourself needing a little extra kick to stand out. Combining noise-free performance with enhanced playing comfort, this sleek instrument is a truly deluxe performer that excels on stage and in the studio.
Three Vintage Noiseless pickups offer all the clarity and articulation you've come to expect from Strat pickups, only with no hum. A push/push mini-switch discretely tucked away between the tone controls, yet still easy to reach, activates the bridge pickup in positions four and five on the selector. Use the bridge and neck pickups together or combine all three pickups for flexible tone that can meet the needs of any song. Enjoy smoother travel when using the tremolo bridge, thanks to the modern two-point synchronized design, which includes bent steel saddles for classic aesthetics. A contoured neck heel and 4-bolt asymmetrical neck plate make it easy to comfortably reach the upper registers of the neck. The flatter 12"-radius fingerboard makes it easy to play fast leads and perform deep bends without fretting out. Locking tuning machines provide enhanced tuning stability, along with quicker string changes.
The ash body bears a highly durable gloss polyurethane finish that will easily stand up to hard playing, keeping the instrument like new for years to come. Classic aesthetic touches include nickel/chrome hardware and a three-ply B/W/B pickguard, along with aged white pickup covers, control knobs and switch tip. Melding classic Fender style and sound with upgraded modern components, the Deluxe Strat is perfect for players who need a wide variety of authentic Fender tones on tap.
- Body Shape - Stratocaster
- Body Material - Ash
- Body Finish - Gloss Polyester
- Neck Material - Maple
- Neck Shape - Modern "C"
- Scale Length - 25.5" (648 mm)
- Fingerboard Radius - 12" (305 mm)
- Number of Frets - 22
- Fret Size - Narrow Tall
- String Nut - Synthetic Bone
- Nut Width - 1.650" (42 mm)
- Truss Rod Nut - 3/16" Hex Adjustment
- Neck Plate - 4-Bolt Asymmetrical
- Neck Finish - Satin Polyurethane
- Fingerboard - Maple
- Position Inlays - Black Pearloid Dots on Maple, White Pearloid Dots on Rosewood
- Bridge Pickup - Vintage Noiseless Single-Coil Strat
- Middle Pickup - Vintage Noiseless Single-Coil Strat
- Neck Pickup - Vintage Noiseless Single-Coil Strat
- Controls - Master Volume, Tone 1. (Neck/Middle Pickups), Tone 2. (Master Tone)
- Pickup Switching - 5-Position Blade: Pos 1. Bridge Pickup Pos 2. Bridge and Middle Pickup Pos 3. Middle Pickup Pos 4. Middle and Neck Pickups (Plus Bridge Pickup When Push/Push Switch Is Activated) Pos 5. Neck Pickup (Plus Bridge Pickup When Push/Push Switch Is Activated)
- Pickup Configuration - SSS
- Bridge - 2-Point Synchronized Tremolo with Bent Steel Saddles
- Tuning Machines - Deluxe Cast/Sealed Locking with Vintage Style Button
- Orientation - Right-Hand
- Pickguard - 3-Ply Mint Green
- Control Knobs - Aged White Plastic
|Number Of Strings
|6 string
|Guitar Style
|stratocaster style
See It In Action
Hear It In Action
Read About It
New: The 2017 Fender American Professional SeriesDecember 2, 2016Read Article
25 Years of Rock'n'Roll: The PMT TimelineSeptember 1, 2016Read Article
PMT Guide To Fender AmpsAugust 26, 2016Read Article
Fender UK Shops - PMT House of RockAugust 11, 2016Read Article
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Relais de Palaiseau
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After 4 days, the RER works
... done. There's something super wonky with the dryer. I'll be drying my clothes and then all of a sudden it just like stops working and all the lights flash. You just have to shut it off and start it over. The other issue is that it sucks. So when it stops working, you have wet clothes, and after you restart it and the cycle finishes, you still have wet clothes. So I have socks drying in my room.
Today I got my email for my immigration appointment with the doctor. At ...
Day 6 and 7: the Eiffel Tower and a country Chatea
... 1300's) to the reign of Louis 14th, he then moved the court to Versailles. It has the tallest medieval keep in Europe at 52 metres tall with a moat and a draw bridge. Love a good castle! The church on site (Saint-Chapelle) was commissioned by Charles the 5th but was completed by Henry 2nd two centuries later. It is reported to contain a relic from Jesus, being a thorn from the Crown of Thorns. The Chateau was originally on an extensive acrerage of park land and forest. Only the ...
Winding along the Seine River
Today is the last day of July, and our last sight-seeing day in Paris. Tomorrow we board a train (bright and early) and head to Craig's final country (I have two more afterwards) of our trip - England! We treated ourselves to another slow morning today, as we have to be up and out of the apartment by 6:15 AM tomorrow morning to catch our train. We didn't have much of a plan for today, which was kind of nice. We had already checked off all of our "must-sees," ...
Last Day...Sacre Couer and Champs Élysées
... by. Fun here. Caught the metro back..paid our hotel bill and made arrangements for a shuttle bus to the airport ..so much easier. AND the elevator has been fixed. Yahoo! Now packing and signing in for tomorrow mornings We are signing off on our blog. I am sure ALL our flights will be on time. We arrive back in Campbell River at 2000 tomorrow June 15. Thank ...
Day One: Paris
... Secondly he has absolutely NO idea how one feels after sitting on a plane for 24 hours, plus some travelling time at each end of the trip. We thanked him for his advice, and when he'd left we quietly went about our unpacking, walked a few blocks to the Supermarket, and came back for a rest. On our way to do some shopping, I bravely asked a nice lady where we could find a Supermarket, and she laughingly told me it ...


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Heavyweight prospect Seth Mitchell talks to Fight Hub TV on his second round win on the Alvarez vs. Hatton undercard. He also shares his views on Alvarez, and the Daniel Ponce De Leon vs. Adrien Bron watch video >>
Seth Mitchell vs Johnathon BanksVacant WBC International heavyweight titleBoardwalk Hall, Atlantic City, New Jersey, USANovember 16, 2012 watch video >>
It's a heavyweight showdown between Seth Mitchell and Chris Arreola. Plus, the return of four-time champion Rafael Marquez to take on Efrain Esquivias. SHOWTIME BOXING: Special Edition, live on Sat watch video >>
For more log onto http://www.fighthubtv.com Like us:http://on.fb.me/xWRC1f follow us:http://bit.ly/wACkIv videos uploaded daily! Subscribe: http://goo.gl/vnzIb Marcos Villegas speaks to Seth Mitchel watch video >>
For more log onto http://www.fighthubtv.comLike us:http://on.fb.me/xWRC1ffollow us:http://bit.ly/wACkIvvideos uploaded daily! Subscribe: http://goo.gl/vnzIbHighlights of the press c watch video >>
¡Prepárate para un doble evento de SHOWTIME BOXING: SPECIAL EDITION, una pelea a 12 asaltos de peso pesado entre Seth "Mayhem" Mitchell y Chris "The Nightmare" Arreola y una batalla a 10 asaltos en p watch video >>
http://fightnetwork.com - Fight Network boxing analyst Corey Erdman previews Seth Mitchell (26-1-1) vs. Chris Arreola (35-3) for the WBC International/WBO NABO heavyweight title at Showtime Boxing Sp watch video >>
Watch as Seth Mitchell, Chris Arreola, Rafael Marquez and Efrain Esquivias hit the scales one day before SHOWTIME BOXING: Special Edition. Watch them live on Saturday, September 7 at 10PM ET/PT on S watch video >>
Chris Arreola vs Seth Mitchell watch video >>


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