Kn. Ehrhard et al., Use of G-protein fusions to monitor integral membrane protein-protein interactions in yeast, NAT BIOTECH, 18(10), 2000, pp. 1075-1079
The control of protein-protein interactions is a fundamental aspect of cell
regulation. Here we describe a new approach to detect the interaction of t
wo proteins in vivo. By this method, one binding partner is an integral mem
brane protein whereas the other is soluble but fused to a G-protein gamma-s
ubunit. If the binding partners interact, G-protein signaling is disrupted.
We demonstrate interaction between known binding partners, syntaxin 1a wit
h 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 la. 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.