Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.
Atypical Recognition Consensus of CIN85/SETA/Ruk SH3 Domains Revealed by Target-assisted Iterative Screening*
Alexei V. Kurakin ,
Susan Wu, and
Dale E. Bredesen
Buck Institute for Age Research, Novato, California 94945
Target-assisted iterative screening applied to random peptidelibraries
unveiled a novel and atypical recognition consensusshared by CIN85/SETA/Ruk
SH3 domains, PX(P/A)XXR. Confirmedby mutagenesis and in
vitro binding experiments, the novelconsensus allowed for the accurate
mapping of CIN85 SH3 bindingsites within known CIN85 interactors, c-Cbl,
BLNK, Cbl-b, AIP1/Alix,SB1, and CD2 proteins, as well as the prediction of
CIN85 novel-interactingpartners in protein databases. Synaptojanin 1, PAK2,
ZO-2,and TAFII70, which contain CIN85 SH3 recognition consensus
sites, were selectively precipitated from mouse brain lysatesby CIN85 SH3
domains in glutathione S-transferase pull-downexperiments. A direct
interaction of synaptojanin 1 and PAK2with CIN85 SH3 domains was confirmed by
Far Western blotting.
Received for publication May 20, 2003.
* This work was supported in part by National Institutes of HealthGrants
NS45093 and AG12282 (to D. E. B.). The costs of publicationof this article
were defrayed in part by the payment of pagecharges. This article must
therefore be hereby marked "advertisement"in accordance
with 18 U.S.C. Section 1734 solely to indicatethis fact.
To whom correspondence should be addressed: Buck Institute for Age Research,
8001 Redwood Blvd., Novato, CA 94945. Tel.: 415-209-22-84; Fax: 415-209-22-30;
The editors suggest the following Related Resources on Science sites:
CIN85 Associates with Multiple Effectors Controlling Intracellular Trafficking of Epidermal Growth Factor Receptors.
K. Kowanetz, K. Husnjak, D. Holler, M. Kowanetz, P. Soubeyran, D. Hirsch, M. H.H Schmidt, K. Pavelic, P. De Camilli, P. A. Randazzo, et al. (2004)
Mol. Biol. Cell
|Abstract »|Full Text »|PDF »
Finding Partners: Emerging Protein Interaction Technologies Applied to Signaling Networks.