Atypical recognition consensus of CIN85/SETA/Ruk SH3 domains revealed by target-assisted iterative screening

被引:49
作者
Kurakin, AV [1 ]
Wu, S [1 ]
Bredesen, DE [1 ]
机构
[1] Buck Inst Age Res, Novato, CA 94945 USA
关键词
D O I
10.1074/jbc.M305264200
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
Target-assisted iterative screening applied to random peptide libraries unveiled a novel and atypical recognition consensus shared by CIN85/SETA/Ruk SH3 domains, PX(P/A) XXR. Confirmed by mutagenesis and in vitro binding experiments, the novel consensus allowed for the accurate mapping of CIN85 SH3 binding sites within known CIN85 interactors, c-Cbl, BLNK, Cbl-b, AIP1/Alix, SB1, and CD2 proteins, as well as the prediction of CIN85 novel-interacting partners in protein databases. Synaptojanin 1, PAK2, ZO-2, and TAF(II)70, which contain CIN85 SH3 recognition consensus sites, were selectively precipitated from mouse brain lysates by CIN85 SH3 domains in glutathione S-transferase pull-down experiments. A direct interaction of synaptojanin 1 and PAK2 with CIN85 SH3 domains was confirmed by Far Western blotting.
引用
收藏
页码:34102 / 34109
页数:8
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