Reporter Epitope Technique for Determining the TransmembraneOrientation of a Membrane Protein Polypeptide Chain
{'Code': '08090200', 'Directorate': {'Abbreviation': 'BIO', 'LongName': 'Direct For Biological Sciences'}, 'Division': {'Abbreviation': 'IOS', 'LongName': 'Division Of Integrative Organismal Systems'}}
{'SignBlockName': 'Stacy Springer', 'PO_EMAI': None, 'PO_PHON': None}
To understand how a membrane protein, such as the cholinergic                   works, it is important to know which parts of its polypeptide                   chain are oriented across the membrane.  The best way to do this                would be x-ray crystallography, but this has not yet been                       possible for membrane proteins as large as nicotinic receptors.                 Sequencing of cDNA clones for membrane protein subunits provides                a great deal of structural information.                                                                                                                         One approach to determining the transmembrane orientation of a                  polypeptide chain is to make antibodies to synthetic peptides                   with the sequence of an interesting part of the polypeptide chain               thought to be exposed on one side of the membrane or the other.                 Then, if these antipeptide antibodies are highly specific, and if               they recognize the native protein, determining to which side of                 the membrane antibodies bind determines the transmembrane                       orientation of that sequence.  The Dr. Lindstrom has developed                  libraries of monoclonal antibodies to nicotinic acetylcholine                   receptors from fish electric organs.  The amino acid sequences to               which some of these antibodies bind, their epitopes, have been                  precisely determined.                                                                                                                                           This research project will use monoclonal antibodies to a                       epitope sequence inserted into the membrane protein to determine                the transmembrane orientation of the epitope when the modified                  DNA is expressed into Xenopus oocytes.  The method will allow                   Dr. Lindstrom to study nicotinic acetylcholine receptors from                   human muscle and chicken and human brain.  If the technique is                  successful, it should allow rapid determination of the                          transmembrane orientation of parts of the polypeptide chain of                  any membrane protein for which the cDNA is available without the                cost, time, effort, and risk of failure of synthesizing peptides,               making antibodies to the peptides, and determining their                        specificity, before testing with the native protein.
{'FirstName': 'Jon', 'LastName': 'Lindstrom', 'PI_MID_INIT': 'M', 'PI_SUFX_NAME': None, 'PI_FULL_NAME': 'Jon M Lindstrom', 'EmailAddress': None, 'NSF_ID': '000311266', 'StartDate': '02/01/1990', 'EndDate': None, 'RoleCode': 'Principal Investigator'}
{'Name': 'The Salk Institute for Biological Studies', 'CityName': 'LA JOLLA', 'ZipCode': '920371002', 'PhoneNumber': '8584534100', 'StreetAddress': '10010 N TORREY PINES RD', 'StreetAddress2': None, 'CountryName': 'United States', 'StateName': 'California', 'StateCode': 'CA', 'CONGRESSDISTRICT': '50', 'CONGRESS_DISTRICT_ORG': 'CA50', 'ORG_UEI_NUM': 'NNJ6BMBTFGN5', 'ORG_LGL_BUS_NAME': 'SALK INSTITUTE FOR BIOLOGICAL STUDIES, SAN DIEGO, CALIFORNIA, THE', 'ORG_PRNT_UEI_NUM': None}
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{'Code': '119200', 'Text': 'NEURONAL AND GLIAL MECHANISMS'}
{'url': 'https://www.nsf.gov/awardsearch/download?DownloadFileName=1988&All=true', 'xml': '8819911.xml'}