CrkL is an adapter for Wiskott-Aldrich syndrome protein and Syk

Atsushi Oda, Hans D. Ochs, Laurence A. Lasky, Susan Spencer, Katsutoshi Ozaki, Mitsuhiro Fujihara, Makoto Handa, Kenji Ikebuchi, Hisami Ikeda

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Wiskott-Aldrich syndrome (WAS) and X-linked thrombocytopenia are caused by mutations of the WAS protein (WASP) gene. WASP may be involved in the regulation of podosome, an actin-rich dynamic cell adhesion structure formed by various types of cells. The molecular links between WASP and podosomes or other cell adhesion structures are unknown. Platelets express an SH2-SH3 adapter molecule, CrkL, that can directly associate with paxillin, which is localized in podosomes. The hypothesis that CrkL binds to WASP was, therefore, tested. Results from coprecipitation experiments using anti-CrkL and GST-fusion proteins suggest that CrkL binds to WASP through its SH3 domain and that the binding was not affected by WASP tyrosine phosphorylation. The binding of GST-fusion SH3 domain of PSTPIP1 in vitro was also not affected by WASP tyrosine phosphorylation, suggesting that the binding of the SH3 domains to WASP is not inhibited by tyrosine phosphorylation of WASP. Anti-CrkL also coprecipitates a 72-kd protein, which was identified as syk tyrosine kinase, critical for collagen induced-platelet activation. CrkL immunoprecipitates contain kinase-active syk, as evidenced by an in vitro kinase assay. Coprecipitation experiments using GST-fusion CrkL proteins suggest that both SH2 and SH3 domains of CrkL are involved in the binding of CrkL to syk. WASP, CrkL, syk, and paxillin-like Hic-5 incorporated to platelet cytoskeleton after platelet aggregation. Thus, CrkL is a novel molecular adapter for WASP and syk and may potentially transfer these molecules to the cytoskeleton through association with cytoskeletal proteins such as Hic-5.

Original languageEnglish
Pages (from-to)2633-2639
Number of pages7
JournalBlood
Volume97
Issue number9
DOIs
Publication statusPublished - 2001 May 1

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

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    Oda, A., Ochs, H. D., Lasky, L. A., Spencer, S., Ozaki, K., Fujihara, M., Handa, M., Ikebuchi, K., & Ikeda, H. (2001). CrkL is an adapter for Wiskott-Aldrich syndrome protein and Syk. Blood, 97(9), 2633-2639. https://doi.org/10.1182/blood.V97.9.2633