| Title: | Use of G-protein fusions to monitor integral membrane protein-protein interactions in yeast |
| Author(s): | Ehrhard KN; Jacoby JJ; Fu XY; Jahn R; Dohlman HG; |
| Address: | "Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06536, USA" |
| ISSN/ISBN: | 1087-0156 (Print) 1087-0156 (Linking) |
| Abstract: | "The control of protein-protein interactions is a fundamental aspect of cell regulation. Here we describe a new approach to detect the interaction of two proteins in vivo. By this method, one binding partner is an integral membrane protein whereas the other is soluble but fused to a G-protein gamma-subunit. If the binding partners interact, G-protein signaling is disrupted. We demonstrate interaction between known binding partners, syntaxin 1a with neuronal Sec1 (nSec1), and the fibroblast-derived growth factor receptor 3 (FGFR3) with SNT-1. In addition, we describe a genetic screen to identify nSec1 mutants that are expressed normally, but are no longer able to bind to syntaxin 1a. This provides a convenient method to study interactions of integral membrane proteins, a class of molecules that has been difficult to study by existing biochemical or genetic methods" |
| Keywords: | "Adaptor Proteins, Signal Transducing Antigens, Surface/genetics/metabolism Cell Membrane/chemistry/*metabolism Cytoplasm/chemistry/metabolism *GTP-Binding Protein gamma Subunits Genes, Reporter/genetics Heterotrimeric GTP-Binding Proteins/*genetics/*metab;" |
| Notes: | "MedlineEhrhard, K N Jacoby, J J Fu, X Y Jahn, R Dohlman, H G eng R01 GM055316/GM/NIGMS NIH HHS/ R01 GM059167/GM/NIGMS NIH HHS/ GM59167/GM/NIGMS NIH HHS/ GM55316/GM/NIGMS NIH HHS/ T32-GM07324/GM/NIGMS NIH HHS/ Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. 2000/10/04 Nat Biotechnol. 2000 Oct; 18(10):1075-9. doi: 10.1038/80274" |
