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J. Biol. Chem., Vol. 281, Issue 28, 19196-19203, July 14, 2006

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Identification of a Novel Inhibitory Actin-capping Protein Binding Motif in CD2-associated Protein^*

Serawit Bruck, Tobias B. Huber, Robert J. Ingham^||, Kyoungtae Kim^¶, Hanspeter Niederstrasser^¶, Paul M. Allen, Tony Pawson^||, John A. Cooper^¶, and Andrey S. Shaw¹

From the Department of Pathology and Immunology, Graduate Program in Molecular Biophysics, ^¶Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110 and ^||Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario MSG 1X5, Canada

CD2-associated protein (CD2AP) is a scaffold molecule that plays a critical role in the maintenance of the kidney filtration barrier. Little, however, is understood about its mechanism of function. We used mass spectrometry to identify CD2AP-interacting proteins. Many of the proteins that we identified suggest a role for CD2AP in endocytosis and actin regulation. To address the role of CD2AP in regulation of the actin cytoskeleton, we focused on characterizing the interaction of CD2AP with actin-capping protein CP. We identified a novel binding motif LXHXTXXRPK(X)₆P present in CD2AP that is also found in its homolog Cin85 and other capping protein-associated proteins such as CARMIL and CKIP-1. CD2AP inhibits the function of capping protein in vitro. Therefore, our results support a role of CD2AP in the regulation of the actin cytoskeleton.

Received for publication, January 6, 2006 , and in revised form, May 11, 2006.

^* This work was supported by National Institutes of Health Grant DK066428-AS and Biophysics Training Grant T32 GM08492. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: Dept. of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110. Tel.: 314-362-4614; Fax: 314-747-4888; E-mail: shaw{at}pathology.wustl.edu.

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