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A novel strategy to identify proteins that interact with the M3 muscarinic receptor in vivo using the split‐ubiquitin system in yeast
Author(s) -
Rosemond Erica,
Li Bo,
McMillin Sally,
Wess Jurgen
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a424-a
Subject(s) - g protein coupled receptor , biology , muscarinic acetylcholine receptor , receptor , tetraspanin , microbiology and biotechnology , muscarinic acetylcholine receptor m5 , rhodopsin like receptors , membrane protein , function (biology) , muscarinic acetylcholine receptor m3 , computational biology , biochemistry , metabotropic receptor , cell , glutamate receptor , membrane
The five muscarinic acetylcholine receptors (M 1 ‐M 5 ) are widely expressed throughout the central and peripheral nervous systems. These receptors are prototypic members of the superfamily of G protein‐coupled receptors (GPCRs). Recent evidence suggests that the function of many GPCRs can be modulated by different classes of receptor‐interacting proteins. The objective of the present study was to identify proteins that can interact with the M 3 muscarinic receptor, a subtype that is found in most regions of the CNS. Recently, the split‐ubiquitin yeast two‐hybrid system has emerged as a novel approach to study membrane protein interactions. Using this novel system, we screened a human brain cDNA library, which led to the identification of 69 proteins that seem to be able to interact with the M 3 receptor in yeast. Most of these proteins have not been previously identified to interact with the M 3 receptor. Interestingly, the candidates include other GPCRs and membrane proteins, several members of the tetraspanin family, various signaling molecules, proteolipid protein‐domain containing proteins and recently cloned proteins of unknown function. It is expected that the results from this screen will provide new insight into the activation, regulation, and/or the physiological function of the M 3 muscarinic receptor. This work was supported by the Intramural Research Program of the NIDDK, NIH.