Constructing glucagon like peptide-1 receptor fused with derivatives of GFP for visualizing protein-protein interaction in living cells.
The glucagon-like peptide-1 receptor (GLP-1 receptor) mediates important antidiabetogenic effects on peripheral tissues. It appears to be one of the most promising therapeutic targets for treatment of diabetes mellitus type 2. Surprisingly, very little is known about the cellular mechanisms that regulate receptor function in living cells. One of the approaches how to study receptor dynamics is by using tagged fluorescent proteins. In this study, YFP-tagged GLP-1 (YFP-GLP-1) receptor and CFP-tagged GLP-1 (CFP-GLP-1) receptor for visualizing protein-protein interaction in living cells were constructed and localized in CHO cells. Cells expressing YFP-GLP-1 and CFP-GLP-1 receptor showed characteristic GLP-1 mediated increase in cAMP, similar to cells expressing a wild type GLP-1 receptor. This means that both types of receptors are functional and localized in plasma membrane.
Bavec
Aljosa
A
Institute of Biochemistry, Faculty of Medicine, University of Ljubljana, Vrazov trg 2, 1000 Ljubljana, Slovenia. aljosa.bavec@mf.uni-lj.si
D000818:Animals; D016466:CHO Cells; D002470:Cell Survival / Q000187:drug effects; D005576:Colforsin / Q000494:pharmacology; D006224:Cricetinae; D003412:Cricetulus; D000242:Cyclic AMP / Q000096:biosynthesis; D052216:Glucagon-Like Peptide 1 / Q000494:pharmacology; D000067757:Glucagon-Like Peptide-1 Receptor; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D006801:Humans; D025941:Protein Interaction Mapping / Q000379:methods*; D021381:Protein Transport / Q000187:drug effects; D018027:Receptors, Glucagon / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*
C000593833:GLP1R protein, human; D000067757:Glucagon-Like Peptide-1 Receptor; D018027:Receptors, Glucagon; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005576:Colforsin; D052216:Glucagon-Like Peptide 1; D000242:Cyclic AMP