A PROTEIN THAT IS HIGHLY RELATED TO GTPASE-ACTIVATING PROTEIN-ASSOCIATED P62 COMPLEXES WITH PHOSPHOLIPASE C-GAMMA

被引:42
作者
MAA, MC
LEU, TH
TRANDEL, BJ
CHANG, JH
PARSONS, SJ
机构
[1] UNIV VIRGINIA, HLTH SCI CTR, CTR CANC, DEPT MICROBIOL, CHARLOTTESVILLE, VA 22908 USA
[2] UNIV VIRGINIA, HLTH SCI CTR, INST MOLEC BIOL, CHARLOTTESVILLE, VA 22908 USA
关键词
D O I
10.1128/MCB.14.8.5466
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
p62 is a highly tyrosyl phosphorylated protein that was first identified in immunoprecipitates of the GTPase-activating protein (GAP) of p21(ras) from cells transformed by oncogenic nonreceptor tyrosine kinases or stimulated through tyrosine kinase receptors (C. Ellis, M. Moran, F. McCormick, and T. Pawson, Nature 343:377-381, 1991). In this article we describe a highly related 62-kDa protein that becomes tyrosyl phosphorylated and associated with phospholipase C gamma (PLC gamma) in C3H10T1/2 cells stimulated with epidermal growth factor (EGF) or transformed by v-src. GAP-associated and PLC gamma-associated p62 comigrated in one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and exhibited nearly identical phosphotryptic peptide patterns. That the association of p62 with PLC gamma was direct and not mediated through binding of GAP-p62 to PLC gamma or to the EGF receptor (and coprecipitation of the receptor with PLC gamma) was demonstrated by (i) the inability to detect GAP in PLC gamma immunocomplexes or PLC gamma in GAP immunocomplexes, (ii) the association of p62 with PLC gamma in v-src-transformed cells in the absence of EGF stimulation, and (iii) in vitro solution binding and direct blotting of p62 with a glutathione S-transferase fusion protein containing the Src homology 2 (SH2) domains of PLC gamma. Unlike GAP, whose N-terminal SH2 mediates the interaction between GAP and p62, PLC gamma was found to require both its N- and C-terminal SH2 regions for p62 binding. These studies demonstrate that a protein identical to or highly related to GAP-associated p62 binds PLC gamma and suggest a means by which ''cross-talk'' between PLC gamma- and GAP-mediated signalling may occur.
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页码:5466 / 5473
页数:8
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