CrkL directs ASAP1 to peripheral focal adhesions

Atsushi Oda, Ikuo Wada, Koichi Miura, Katsuya Okawa, Toshihiko Kadoya, Takashi Kato, Hiroshi Nishihara, Masae Maeda, Shinya Tanaka, Kazuo Nagashima, Chiaki Nishitani, Kazuhiko Matsuno, Masaho Ishino, Laura M. Machesky, Hiroyoshi Fujita, Paul Randazzo

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    Abstract

    Searching for proteins in platelets that can interact with the N-terminal SH3 domain of CrkL (using a combination of a pull-down assay followed by mass spectrometry), we have found that human platelets express an ADP-ribosylation factor (Arf)-specific GTPase-activating protein (GAP), ASAP1, as a CrkL-binding protein. In spreading platelets, most endogenous ASAP1 is localized at peripheral focal adhesions. To determine the physiologic significance of the CrkL-ASAP1 association, we overexpressed CrkL, ASAP1, or both in combination in COS7 cells. Unlike endogenous ASAP1 in platelets, overexpressed ASAP1 showed diffuse cytoplasmic distribution. However, when co-expressed with wild-type CrkL, both endogenous and expressed ASAP1 accumulated at CrkL-induced focal adhesions. An SH2-mutated CrkL, which cannot localize at focal adhesions, failed to recruit ASAP1 into focal adhesions. Thus, CrkL appears to be a lynchpin between ASAP1 and peripheral focal adhesions.

    Original languageEnglish
    Pages (from-to)6456-6460
    Number of pages5
    JournalJournal of Biological Chemistry
    Volume278
    Issue number8
    DOIs
    Publication statusPublished - 2003 Feb 21

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    ASJC Scopus subject areas

    • Biochemistry

    Cite this

    Oda, A., Wada, I., Miura, K., Okawa, K., Kadoya, T., Kato, T., Nishihara, H., Maeda, M., Tanaka, S., Nagashima, K., Nishitani, C., Matsuno, K., Ishino, M., Machesky, L. M., Fujita, H., & Randazzo, P. (2003). CrkL directs ASAP1 to peripheral focal adhesions. Journal of Biological Chemistry, 278(8), 6456-6460. https://doi.org/10.1074/jbc.M210817200