Xk. Tong et al., The endocytic protein intersectin is a major binding partner for the Ras exchange factor mSos1 in rat brain, EMBO J, 19(6), 2000, pp. 1263-1271
We recently identified intersectin, a protein containing two EH and five SH
3 domains, as a component of the endocytic machinery. The N-terminal SH3 do
main (SH3A), unlike other SH3 domains from intersectin or various endocytic
proteins, specifically inhibits intermediate events leading to the formati
on of clathrin-coated pits. We have now identified a brain-enriched, 170 kD
a protein (p170) that interacts specifically with SH3A. Screening of combin
atorial peptides reveals the optimal ligand for SH3A as Pp(V/I)PPR, and the
170 kDa mammalian son-of-sevenless (mSos1) protein, a guanine-nucleotide e
xchange factor for Ras, contains two copies of the matching sequence, PPVPP
R. Immunodepletion studies confirm that p170 is mSos1. Intersectin and mSos
1 are co-enriched in nerve terminals and are co-immunoprecipitated from bra
in extracts. SH3A competes with the SH3 domains of Grb2 in binding to mSos1
, and the intersectin-mSos1 complex can be separated from Grb2 by sucrose g
radient centrifugation, Overexpression of the SH3 domains of intersectin bl
ocks epidermal growth factor-mediated Ras activation. These results suggest
that intersectin functions in cell signaling in addition to its role in en
docytosis and may link these cellular processes.