Tyrosine phosphorylated Disabled 1 recruits Crk family adapter proteins.
Disabled 1 (Dab1) functions as a critical adapter protein in the Reelin signaling pathway to direct proper positioning of neurons during brain development. Reelin stimulates phosphorylation of Dab1 on tyrosines 198 and 220, and phosphorylated Dab1 is likely to interact with downstream signaling proteins that contain Src homology 2 (SH2) domains. To search for such proteins, we used a Sepharose-conjugated peptide containing phosphotyrosine 220 (PTyr-220) of Dab1, as an affinity matrix to capture binding proteins from mouse brain extracts. Mass spectrometric analysis of bound proteins revealed that Crk family adapter proteins selectively associated with this phosphorylation site. We further show that Crk-I and Crk-II, but not CrkL, stimulate phosphorylation of Dab1 on tyrosine 220 in a Src-dependent manner. Our results suggest that Crk family adapter proteins may play an important role in the Reelin signaling pathway during brain development.
Huang
Yongcheng
Y
Division of Pharmacology, College of Pharmacy, The Ohio State University, 500 W. 12th Avenue, Columbus, OH 43210, USA.
Magdaleno
Susan
S
Hopkins
Rachel
R
Slaughter
Clive
C
Curran
Tom
T
Keshvara
Lakhu
L
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D001921:Brain / Q000196:embryology / Q000378:metabolism; D002460:Cell Line; D015870:Gene Expression; D006801:Humans; D051379:Mice; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D050737:Proto-Oncogene Proteins c-crk; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D000091042:Reelin Protein; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D014162:Transfection; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism*; D018909:src Homology Domains; D019061:src-Family Kinases / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; C115785:DAB1 protein, human; D009419:Nerve Tissue Proteins; D011518:Proto-Oncogene Proteins; D050737:Proto-Oncogene Proteins c-crk; D011994:Recombinant Proteins; D000091042:Reelin Protein; D014443:Tyrosine; D019061:src-Family Kinases; C000717127:RELN protein, human; C000717128:Reln protein, mouse