interacts_with DB00904?
Chronic methamphetamine exposure alters immune function in normal and retrovirus-infected mice . Methamphetamine ( MA ) abuse represents a growing problem in the USA with an increase of sudden death . To evaluate the immune function alterations due to chronic methamphetamine use , we examined C57BL/C mice with LP-BM5 retrovirus infection plus methamphetamine exposure . Mice were randomly assigned to the following groups : placebo , placebo retrovirus-infected , uninfected MA treated and retrovirus-infected MA treated . Placebo , MA-treated groups were intraperitoneally injected with saline , MA , respectively , with a gradually increasing dose from 15 to 40 mg/kg for 12 weeks ( 5 days/week ) . Con A- and LPS-induced mitogenesis of splenocytes , cytokine production by splenocytes culture and lipid peroxides in the liver were measured . Heart tissue histopathology was analyzed in all the groups with murine cytomegalovirus ( CMV ) superinfection . Our data showed that MA treatment significantly decreased production of P60568 and interferon gamma ( P01579 ) in uninfected mice but did not further suppress the reduced Th1 cytokines in retrovirus-infected mice . There were no significant effects on cytokines P05112 and P05231 . However , tumor necrosis factor ( P01375 ) was significantly increased in both uninfected and infected mice due to MA treatment . Lipid peroxides in liver were significantly increased both in uninfected and retrovirus-infected mice due to MA exposure . DB00163 levels in liver were significantly decreased in uninfected mice due to MA treatment . CMV superinfection greatly increased the cardiac lesions in retrovirus-infected mice while no significant histopathology changes were detected due to MA treatment . Our data suggest that MA has immunomodulation activity , suppressing Th1 cytokine production and enhancing some Th2 cytokine secretion , as well as increasing lipid peroxides in uninfected mice . The interaction between LP-BM5 and MA remains unclear . Diabetes mellitus in cancer patients treated with combination interleukin 2 and alpha-interferon . Diabetes mellitus is thought to be an autoimmune disease caused by destruction of beta cells in pancreatic islets . P01308 resistance in the peripheral tissues may also play a role . Both interleukin 2 ( P60568 ) and alpha interferon can enhance immune function by stimulating formation of cytolytic T cells and/or antigen expression on both normal and tumor cells . This report describes three patients with advanced malignancy who were treated with combination P60568 and alpha interferon who had the onset or worsening of diabetes mellitus . One patient died as a result . There is evidence that interferon can increase insulin resistance and it is likely that both agents can initiate or enhance an ongoing autoimmune process . Physicians using this combination of drugs should be aware of this potential serious toxicity . Hsp27 regulates epithelial mesenchymal transition , metastasis , and circulating tumor cells in prostate cancer . Defining the mechanisms underlying metastatic progression of prostate cancer may lead to insights into how to decrease morbidity and mortality in this disease . An important determinant of metastasis is epithelial-to-mesenchymal transition ( EMT ) , and the mechanisms that control the process of EMT in cancer cells are still emerging . Here , we report that the molecular chaperone Hsp27 ( P04792 ) drives EMT in prostate cancer , whereas its attenuation reverses EMT and decreases cell migration , invasion , and matrix metalloproteinase activity . Mechanistically , silencing Hsp27 decreased P05231 -dependent P40763 phosphorylation , nuclear translocation , and P40763 binding to the Twist promoter , suggesting that Hsp27 is required for P05231 -mediated EMT via modulation of P40763 /Twist signaling . We observed a correlation between Hsp27 and Twist in patients with prostate cancer , with Hsp27 and Twist expression each elevated in high-grade prostate cancer tumors . Hsp27 inhibition by DB06094 , an antisense therapy currently in phase II trials , reduced tumor metastasis in a murine model of prostate cancer . More importantly , DB06094 treatment decreased the number of circulating tumor cells in patients with metastatic castration-resistant prostate cancer in a phase I clinical trial . Overall , this study defines Hsp27 as a critical regulator of P05231 -dependent and P05231 -independent EMT , validating this chaperone as a therapeutic target to treat metastatic prostate cancer . P01308 -like growth factor-I is a serum component stimulating growth of human neuroblastoma . Human non-autocrine neuroblastoma cells SK-N-SH and LF require serum for proliferation in vitro . We wished to determine the role of serum-borne insulin-like growth factor I ( P05019 ) as mitogen for these cells . Introduction of the monoclonal antibody alpha-IR3 against human P08069 reduced proliferation in the presence of fetal bovine serum ( FBS ) . P05019 ( 5 nM ) was as effective as FBS ( 10 % ) in stimulating proliferation . DB00071 mimicked the effects of P05019 , but at a 1000-fold higher concentration . The antibody alpha-IR3 reduced growth stimulated by P05019 more effectively than growth stimulated by insulin . Thus , proliferation of human non-autocrine neuroblastoma cells can be effectively manipulated by exogenous P05019 . A cytokine-controlled mechanism for integrated regulation of T-lymphocyte motility , adhesion and activation . The co-ordination of T-cell motility , adhesion and activation remains poorly understood . It is also unclear how these functions are co-ordinated with external stimuli . Here we unveil a series of molecular interactions in cis at the surface of T lymphocytes with potent effects on motility and adhesion in these cells , and communicating with proliferative responses . These interactions were controlled by the signature cytokines of T helper subsets interleukin-2 ( P60568 ) and P05112 . P01130 -related protein 1 ( Q07954 ) was found to play a key role for T-cell motility by promoting development of polarized cell shape and cell movement . Endogenous thrombospondin-1 ( P07996 -1 ) enhanced cell surface expression of Q07954 through Q08722 . Cell surface expressed Q07954 induced motility and processing of P07996 -1 while inhibiting adhesion to intercellular adhesion molecule 1 and fibronectin . P60568 , but not P05112 , stimulated synthesis of P07996 -1 and motility through P07996 -1 and Q07954 . Stimulation of the T-cell receptor ( TCR ) /CD3 complex inhibited P07996 -1 expression . Inhibitor studies indicated that Q07954 regulated P07996 -1 expression and promoted motility through JAK signalling . This Q07954 -mediated motogenic signalling was connected to Q08722 /Gi protein signalling and P60568 -induced signalling through P07996 -1 . The motogenic P07996 -1/ Q07954 mechanism antagonized TCR/CD3-induced T-cell proliferation . These results indicate that Q07954 in collaboration with P07996 -1 directs a counter-adhesive and counter-proliferative motogenic cascade . T cells seem programmed to prioritize movement before adhesion through this cascade . In conclusion , vital decision-making in T lymphocytes regulating motility , adhesive interactions and proliferation , are integrated through a molecular mechanism connecting different cell surface receptors and their signalling pathways . The Lyme disease vaccine takes its toll . Vaccination with Borrelia burgdorferi outer surface protein ( Osp ) A can induce a protective response against Lyme disease and serves as a model to understand the generation of protective immune responses against the spirochete . The innate response to pathogens is activated by specific Toll-like receptors ( TLRs ) that recognize distinct pathogen-associated molecular patterns . O60603 is of particular interest for B. burgdorferi research because O60603 recognizes several pathogen-associated molecular patterns , including lipoproteins . O60603 may form heterodimers with Q9Y2C9 to identify diacylated lipoproteins , while O60603 works in concert with Q15399 to recognize triacylated lipoproteins such as OspA . We will discuss the role of Q15399 /2 in the development of responses to OspA in Q15399 -and O60603 -deficient mice , and in selected individuals that received the OspA vaccine . While > 95 % of human OspA-based Lyme disease vaccine recipients develop OspA antibodies , a very small group of individuals did not develop detectable humoral responses against OspA . We demonstrated that this hyporesponsiveness to OspA vaccination was associated with decreased cell surface expression of Q15399 . Moreover , Q15399 - and O60603 -deficient mice did not develop significant levels of OspA antibodies following vaccination with OspA , providing a correlation with human hyporesponsiveness to OspA . These data suggest that defects in the Q15399 /2 signaling pathway are associated with an impaired ability to generate antibodies following immunization with DB00045 . Comparison of the novel antipsychotic ziprasidone with clozapine and olanzapine : inhibition of dorsal raphe cell firing and the role of P08908 receptor activation . Ziprasidone is a novel antipsychotic agent which binds with high affinity to P08908 receptors ( Ki = 3.4 nM ) , in addition to P28221 , 5-HT2 , and D2 sites . While it is an antagonist at these latter receptors , ziprasidone behaves as a P08908 agonist in vitro in adenylate cyclase measurements . The goal of the present study was to examine the P08908 properties of ziprasidone in vivo using as a marker of central P08908 activity the inhibition of firing of serotonin-containing neurons in the dorsal raphe nucleus . In anesthetized rats , ziprasidone dose-dependently slowed raphe unit activity ( ED50 = 300 micrograms/kg i.v. ) as did the atypical antipsychotics clozapine ( ED50 = 250 micrograms/kg i.v. ) and olanzapine ( ED50 = 1000 micrograms/kg i.v. ) . Pretreatment with the P08908 antagonist WAY-100,635 ( 10 micrograms/kg i.v. ) prevented the ziprasidone-induced inhibition ; the same dose of WAY-100,635 had little effect on the inhibition produced by clozapine and olanzapine . Because all three agents also bind to alpha 1 receptors , antagonists of which inhibit serotonin neuronal firing , this aspect of their pharmacology was assessed with desipramine ( DB01151 ) , a NE re-uptake blocker previously shown to reverse the effects of alpha 1 antagonists on raphe unit activity . DB01151 ( 5 mg/kg i.v. ) failed to reverse the inhibitory effect of ziprasidone but produced nearly complete reversal of that of clozapine and olanzapine . These profiles suggest a mechanism of action for each agent , P08908 agonism for ziprasidone and alpha 1 antagonism for clozapine and olanzapine . The P08908 agonist activity reported here clearly distinguishes ziprasidone from currently available antipsychotic agents and suggests that this property may play a significant role in its pharmacologic actions . Comparison of the antiemetic effects of a P08908 agonist , LY228729 , and 5- Q9H205 antagonists in the pigeon . Vomiting may be induced by a variety of agents such as drugs , oncolytics , and provocative motion , as well as being conditioned to occur to environmental stimuli . Such emesis has recently been shown to be blocked by agonists at the P08908 subtype of serotonin receptor . The antiemetic effects of LY228729 [ (-)-4-(dipropylamine)-1,3,4,5-tetrahydrobenz-(c,d)indole-6- carboxamide ] , a P08908 receptor agonist , were tested and compared to the antiemetic effects of the 5- Q9H205 receptor antagonists ondansetron , tropisetron , and MDL 72222 ( 3-tropanyl-3,5-dichlorobenzoate ) . The emetic stimuli tested are known to be blocked by 5- Q9H205 antagonists in species other than the pigeon . In the pigeon , LY228729 totally abolished vomiting induced by fully emetic doses of cisplatin ( 10 mg/kg ) , ipecac ( 3 ml/kg ) , emetine ( 10 mg/kg ) , and a 5- Q9H205 agonist , m-(chlorophenyl)-biguanide ( 1.25 mg/kg ) . MDL 72222 blocked ipecac-induced vomiting in a dose-related manner and was partially effective in attenuating cisplatin-induced emesis . DB00904 and tropisetron were partially effective in blocking emetine- and mCPBG-induced vomiting . DB00904 exhibited an intrinsic emetic response that could not be blocked by MDL 7222 , but which was eliminated by LY228729 . It was concluded that P08908 agonists are more effective in the pigeon than are 5- Q9H205 antagonists against these types of emetic stimuli . These results broaden the range of emetic stimuli that are blocked by P08908 agonists in the pigeon . Influence of immunomodulatory drugs on the cytotoxicity induced by monoclonal antibody 17-1A and interleukin-2 . Patients treated with monoclonal antibodies and cytokines for cancer receive often co-medication , which may influence treatment efficacy . Therefore , we investigated with a flowcytometric cytotoxicity assay the effect of several immunomodulatory drugs on antibody dependent cellular cytotoxicity ( ADCC ) , interleukin-2 ( P60568 ) induced cytotoxicity and P60568 -induced-ADCC . We found that dexamethasone markedly inhibited the P60568 induced cytotoxicity and the P60568 -induced-ADCC . DB00904 , a P46098 serotonin receptor antagonist augmented significantly ADCC . Clemastine , a histamine type-2 receptor antagonist augmented the P60568 -induced-ADCC . The P01375 antagonist thalidomide suppressed ADCC whereas pentoxifylline proved to be ineffective . Other tested drugs namely ibuprofen and indomethacin , both prostaglandin E2 antagonists , cimetidine a histamine type-2 receptor antagonist , the opioid pethidine , prostaglandin E2 and histamine exerted minor effects or had no influence on the tested parameters . We conclude that glucocorticosteroids should be avoided with monoclonal antibody and cytokine treatment . According to our in vitro data the other drugs tested did not have a negative impact on cellular cytotoxicity and ADCC . Raf inhibitor stabilizes receptor for the type I interferon but inhibits its anti-proliferative effects in human malignant melanoma cells . DB05258 ( IFNalpha ) is widely used in treatment of malignant melanoma patients . This cytokine acts on cells by engaging Type I IFN receptor consisting of two subunits , ( P17181 and P48551 ) followed by activation of Janus kinases ( Jak ) . Levels of P17181 ( regulated via degradation mediated by the betaTrcp E3 ubiquitin ligase ) and IFNalpha signaling were reduced in 1205Lu melanoma cell line that harbors activated P15056 and exhibits high levels of betaTrcp ubiquitin ligase . Expression of stabilized P17181 in melanoma cells decreased their tumorigenicity . Furthermore , RNAi-mediated P15056 knockdown and pharmacologic inhibition of either Raf or Q02750 decreased levels of betaTrcp and stabilized P17181 . However , despite causing stabilization of P17181 , Raf inhibitor BAY 43-9006 interfered with cellular responses to IFNalpha most likely due to its ability to directly inhibit Jak activity . We discuss the implications of this result for combination therapy with BAY 43-9006 and IFNalpha in melanoma patients . Human tumor infiltrating lymphocytes . Analysis of lymphokine mRNA expression and relevance to cancer immunotherapy . Tumor infiltrating lymphocytes ( Q15399 ) isolated from 12 patients with metastatic malignant melanoma , renal cell carcinoma , or breast adenocarcinoma were expanded in rIL-2 for 22 to 45 days ( median 33 days ) and analyzed for lymphokine mRNA expression and patterns of TCR gene rearrangement . All Q15399 cultures were significantly enriched for T cells , with CD3+ CD8+ cells predominant in 8 of 10 cases tested , and demonstrated an oligoclonal ( rather than polyclonal ) pattern of TCR gene rearrangement . Nine of 12 cultures could effectively lyse the autologous targets in short term chromium release assays . P60568 expanded- Q15399 expressed mRNA for P01375 and P01374 ( lymphotoxin ) and , in 5 of 9 ( 41 % ) cases , granulocyte/macrophage-colony stimulating factor mRNA but not P01584 or P60568 transcripts . Cultured Q15399 deprived of rIL-2 for 4 days did not constitutively express mRNA for any of the lymphokines tested . One long term Q15399 line in culture was followed and periodically tested for lytic activity and P01375 -mRNA expression . Loss of the specific cytolytic but not proliferative activity at day 85 was associated with disappearance of P01375 mRNA . Profiles of lymphokine secretion may provide a useful marker for functionally characterizing different T cell subsets and may provide correlates of the in vivo anti-tumor effects of these cells when Q15399 are adoptively transferred into cancer-bearing patients . Regulation of apoptosis by tyrosine-containing domains of IL-4R alpha : Y497 and Y713 , but not the P42226 -docking tyrosines , signal protection from apoptosis . P05112 is a cytokine with important antiapoptotic activity . We have analyzed the role that tyrosine-containing domains within the cytoplasmic tail of IL-4R alpha play in P05112 -mediated protection from apoptosis . 32D cells expressing a wt huIL-4R alpha or one truncated at aa 557 were protected by huIL-4 from apoptosis while cells expressing a receptor truncated at aa 657 were not , suggesting that the carboxyl-terminal domain signals protection from apoptosis . However , changing Y713 within this region to phenylalanine had no effect . To analyze the contribution of tyrosine-containing domains independently , we transplanted regions of the huIL-4R alpha to a truncated form of the huIL-2R beta that could not signal protection from apoptosis . Transplantation of the huIL-4R alpha domains containing Y497 or Y713 partially prevented cell death and together signaled protection from apoptosis in response to P60568 as well as the wt IL-2R beta . Mutation of Y497 and Y713 to phenylalanine inhibited protection . In contrast , transplantation of the domain containing the potential P42226 -docking tyrosines alone had no effect , yet it inhibited the protection mediated by the other domains . Although IL-4R alpha signals Shc and SH2-containing inositol phosphatase ( Q92835 ) phosphorylation , we could not establish an association between their activation and protection from apoptosis . Taken together , this study suggests that the domains of the huIL-4R alpha containing Y497 and Y713 positively regulate protection from apoptosis while the domain containing the P42226 docking sites suppresses this protection , and that additional signaling molecules other than insulin receptor substrate-1 ( P35568 ) , Shc , or Q92835 may be involved in antiapoptotic signaling . P55157 inhibitor decreases plasma cholesterol levels in P01130 -deficient WHHL rabbits by lowering the VLDL secretion . To examine whether a microsomal triglyceride transfer protein ( P55157 ) -inhibitor is effective in patients with homozygous familial hypercholesterolemia , we administered ( 2S ) -2-cyclopentyl-2-[4-[(2,4-dimethyl-9H-pyrido[2,3-b]indol-9-yl)methyl]phenyl]-N- [ ( 1S ) -2-hydroxy-1-phenylethyl ] ethanamide ( DB04852 ) , a new P55157 inhibitor , to low-density lipoprotein ( LDL ) -receptor-deficient Watanabe heritable hyperlipidemic ( WHHL ) rabbits at doses of 3 , 6 , and 12 mg/kg for 4 weeks . In the 12 mg/kg group , the plasma cholesterol and triglyceride levels were decreased by 70 % and 45 % , respectively , and the very low-density lipoprotein ( VLDL ) secretion rate was decreased by 80 % . The composition of newly secreted VLDL was similar in each group . This suggests that DB04852 diminished the number of VLDL particles secreted from the liver . Although the ratio of vitamin E/LDL was not altered by DB04852 , triglyceride accumulation and a decrease in vitamin E were observed in the liver . In conclusion , an inhibition of VLDL secretion led to a decrease of plasma LDL in WHHL rabbits , and P55157 inhibitors should have hypolipidemic effects against homozygous familial hypercholesterolemia . Differential antiinflammatory and antinociceptive effects of the somatostatin analogs octreotide and pasireotide in a mouse model of immune-mediated arthritis . OBJECTIVE : Clinical and preclinical evidence suggests that somatostatin exhibits potent antiinflammatory and antinociceptive properties . However , it is not known which of the 5 somatostatin receptor subtypes ( SSTRs 1-5 ) is involved in these actions . The purpose of this study was to assess the effects of the stable somatostatin analogs octreotide and pasireotide ( DB06663 ) in a mouse model of antigen-induced arthritis ( AIA ) . METHODS : Studies were performed in P30874 -deficient mice ( P30874 (-/-) ) and their wild-type littermates ( P30874 (+/+) ) . The expression of P30872 , SSTR2A , P32745 , and P35346 in dorsal root ganglia was examined by immunohistochemistry . RESULTS : Untreated P30874 (-/-) mice with AIA displayed joint swelling and mechanical hyperalgesia similar to that seen in P30874 (+/+) mice . In wild-type mice , both octreotide and pasireotide significantly attenuated knee joint swelling and histopathologic manifestations of arthritis to an extent comparable to that of dexamethasone . In P30874 (-/-) mice , the antiinflammatory effects of both octreotide and pasireotide were completely abrogated . Prolonged administration of pasireotide also inhibited joint swelling and protected against joint destruction during AIA flare reactions . In addition , both octreotide and pasireotide reduced inflammatory hyperalgesia . The antinociceptive actions of octreotide were abolished in P30874 (-/-) mice , but those of pasireotide were retained . In dorsal root ganglia of naive wild-type mice , only P30872 and SSTR2A , but not P32745 or P35346 , were detected in a subset of small- and medium-diameter neurons . CONCLUSION : Our findings indicate that the antinociceptive and antiinflammatory actions of octreotide and pasireotide are largely mediated via the P30874 receptor . In addition , we identified the P30872 receptor as a novel pharmacologic target for somatostatin-mediated peripheral analgesia in inflammatory pain . Synergism of Q9HC29 and Q96P20 activators promotes a unique transcriptional profile in murine dendritic cells . NLRs are cytoplasmic proteins that sense cellular stress and intracellular damage resulting from pathogen uptake . To date , the role of NLRs has been studied using combinations of NLR and TLR agonists , but the interplay between two different NLRs remains uncharacterized . In this study , we employed microarrays to investigate in DCs the regulation of gene transcription mediated by activation of Q9HC29 and Q96P20 pathways using P16444 and MSU . P16444 and MSU co-stimulation of murine BMDCs up-regulated the expression of genes encoding molecules for antigen presentation and co-stimulation ( MHC class II , P33681 , P42081 ) , integrins ( P05106 , P06756 ) , cytokines ( IL-1α , IL-1β , P05231 , P60568 , Q9NPF7 , IL-12p40 ) , and chemokines ( P09341 , P19875 ) . Transcription of the cytokine genes induced by P16444 and MSU partially depended on Q9HC29 but was independent of Q96P20 . Finally , we showed that P27361 and c- P05412 activation increased upon P16444 and MSU co-stimulation . As a whole , the results indicate that two different NLR activators synergize at the transcriptional level , leading to unique differential expression of genes involved in the innate immune response . Effects of chimeric somatostatin-dopamine molecules on human peripheral blood lymphocytes activation . O43521 23A761 , selective for somatostatin receptors subtypes 2 , 5 and the dopamine receptor subtype 2 , and O43521 23A757 with affinity for P30874 and DAR2 were studied on human PBL proliferation and activation . O43521 23A761 was significantly more potent than specific SSTR and DAR2 agonists in suppressing lymphocyte proliferation induced by mitogen or alloantigen , while O43521 23A757 was more potent than specific P30874 and DAR2 agonists in suppressing antigen induced proliferation only . Both molecules displayed enhanced potency in suppressing IFNgamma and P05231 secretion compared with the SSTR and DAR2 analogs , while only O43521 23A761 was able to inhibit P60568 secretion and its effect is more potent than the control analogs . Furthermore O43521 23A761 inhibit cell progression into the S phase and then into the G2/M , while O43521 23A757 inhibited bromodeoxyuridine incorporation only during the S phase . Both chimeric molecules resulted significantly more effective than the respective controls . DB04998 inhibits activation of nuclear factor-kappaB ( NF-kappaB ) by forming a complex with NF-kappaB essential modulator ( Q9Y6K9 ) and nucleolin . DB04998 , also known as DB04998 , is an experimental anticancer drug that recently entered human clinical trials . It is a member of a novel class of antiproliferative agents known as G-rich oligonucleotides ( P09341 ) , which are non-antisense , guanosine-rich phosphodiester oligodeoxynucleotides that form stable G-quadruplex structures . The biological activity of GROs results from their binding to specific cellular proteins as aptamers . One important target protein of GROs has been previously identified as nucleolin , a multifunctional protein expressed at high levels by cancer cells . Here , we report that DB04998 also associates with nuclear factor-kappaB ( NF-kappaB ) essential modulator ( Q9Y6K9 ) , which is a regulatory subunit of the inhibitor of kappaB ( IkappaB ) kinase ( IKK ) complex , and also called IKKgamma . In the classic NF-kappaB pathway , the IKK complex is required for phosphorylation of P25963 and subsequent activation of the transcription factor NF-kappaB . We found that treatment of cancer cells with DB04998 inhibits IKK activity and reduces phosphorylation of P25963 in response to tumor necrosis factor-alpha stimulation . Using a reporter gene assay , we showed that DB04998 blocks both tumor necrosis factor-alpha-induced and constitutive NF-kappaB activity in human cancer cell lines derived from cervical , prostate , breast , and lung carcinomas . In addition , we showed that , in DB04998 -treated cancer cells , Q9Y6K9 is coprecipitated by nucleolin , indicating that both proteins are present in the same complex . Our studies suggest that abrogation of NF-kappaB activity may contribute to the anticancer effects of DB04998 and that nucleolin may play a previously unknown role in regulating the NF-kappaB pathway . Met326Ile aminoacid polymorphism in the human p85 alpha gene has no major impact on early insulin signaling in type 2 diabetes . Class I alpha phosphatidylinositol ( PI ) 3-kinase is an important enzyme in the early insulin signaling cascade , and plays a key role in insulin-mediated glucose transport . Despite extensive investigation , the genes responsible for the development of the common forms of type 2 diabetes remain unknown . This study was performed to identify variants in the coding region of p85 alpha , the regulatory subunit of PI 3-kinase . Fibroblasts from skin biopsies from type 2 diabetics and controls were established to address this issue . P85 alpha cDNA was sequenced , and a single point mutation at codon 326 was found . This mutation resulted in a homozygous missense amino acid change DB00134 --> DB00167 in one subject with type 2 diabetes and heterozygous variant in two other diabetic patients and one with severe insulin resistance . Interestingly , those patients revealed an impaired insulin-mediated insulin receptor substrate ( P41252 ) -1 binding to p85 alpha without any alteration in Q9Y4H2 /p85 alpha association . Furthermore , P35568 , Q9Y4H2 , p85 alpha and MAPK protein contents were not significantly changed , and neither were MAPK or Akt phosphorylation . We conclude from our data that this variant may have only minor impact on signaling events ; however , in combination with variants in other genes encoding signaling proteins , this may have a functional impact on early insulin signaling . Contribution of the Box 1 and Box 2 motifs of cytokine receptors to Jak1 association and activation . Kinases of the Jak family ( Jak1/2/3 and Tyk2 ) interact with the membrane proximal domain of different cytokine receptors and play a critical role in the activation of cytokine and growth factor signaling pathways . In this report we demonstrate that both the Box 1 and Box 2 motif collaborate in the association and activation of Jak1 by type I interferons . Mutational analysis of the beta chain of type I interferon receptor ( IFNalphaRbetaL/ P48551 ) revealed that Box 1 plays a more significant role in activation than in the association with Jak1 . On the contrary , the Box 2 motif contributes more to the association with Jak1 than to kinase activation . Additionally , the study of the Jak1 binding sites on the P60568 receptor beta ( IL2Rbeta ) , IFNgammaRalpha/ P15260 , and IL10Ralpha/IL10R1 chains suggests that cytokine receptors have two different kinds of interaction with Jak1 . One form of interaction involves the Box 1 and the previously described Box 2 motif , which we now designate as Box 2A , characterized by the VEVI and LEVL sequences present in IFNalphaRbetaL/ P48551 and IL2Rbeta subunits , respectively . The second form of interaction requires a motif termed Box 2B , which is present in the IFNgammaRalpha/ P15260 ( SILLPKS ) and IL10Ralpha/IL10R1 ( SVLLFKK ) chains . Interestingly , Box 2B localizes close to the membrane region ( 8-10 amino acids from the membrane ) similar to Box 1 , whereas Box 2A is more distal ( 38-58 amino acids from the membrane ) .