RRC ID 12044
Author Koga S, Yogo K, Yoshikawa K, Samori H, Goto M, Uchida T, Ishida N, Takeya T.
Title Physical and functional association of c-Src and adhesion and degranulation promoting adaptor protein (ADAP) in osteoclastogenesis in vitro.
Journal J Biol Chem
Abstract c-Src plays a crucial role in osteoclastogenesis. In this study, we searched for c-Src-binding proteins using a combination of pull-down assays and mass spectrometric analysis, and identified the association of adhesion and degranulation promoting adaptor protein (ADAP) with c-Src in RAW264 cells and osteoclast precursors prepared from bone marrow cells. The kinase activity and the SH2 domain of c-Src were required for this association and Tyr807 in the extreme carboxyl terminus of ADAP was identified as a major recognition site. ADAP was found to be expressed in cells at the prefusion stage and localized mainly in the leading edge of lamellipodia and in pseudopodia. Tyrosine phosphorylation of ADAP was induced in an integrin-dependent manner, and the level was Src kinase-dependent. ADAP-knockdown RAW264 cells showed retarded migration and formed few multinucleated cells. Cas, known to be phosphorylated by c-Src, was identified as a major tyrosine-phosphorylated protein in differentiating RAW264 cells and the phosphorylation appeared to be decreased in ADAP-knockdown cells. ADAP thus may play an important role as a partner of c-Src for cell migration and progression to the multinucleated cell stage in osteoclastogenesis in vitro.
Volume 280(36)
Pages 31564-71
Published 2005-9-9
DOI 10.1074/jbc.M502703200
PII S0021-9258(20)79270-7
PMID 16020549
MeSH Adaptor Proteins, Signal Transducing / genetics Adaptor Proteins, Signal Transducing / physiology* Animals CSK Tyrosine-Protein Kinase Cell Adhesion / physiology Cell Degranulation / physiology* Cell Differentiation / physiology* Cell Line Cell Movement / physiology Crk-Associated Substrate Protein Female Gene Expression Profiling Humans Mice NIH 3T3 Cells Osteoclasts / cytology Osteoclasts / physiology* Phosphorylation Protein-Tyrosine Kinases / physiology* Proteins / metabolism RNA, Small Interfering / metabolism Retinoblastoma-Like Protein p130 Stem Cells / cytology Stem Cells / physiology Tyrosine / genetics Tyrosine / metabolism src-Family Kinases
IF 4.238
Times Cited 10
DNA material pCAhMCSF Human M-CSF (RDB01524)