Direct regulation of ZAP-70 by SHP-1 in T cell antigen receptor signaling

Science

Volumn 272, Issue 5265, 1996, Pages 1173-1176

  Plas, David R ^a   Johnson, Robin ^a   Pingel, Jeanette T ^a   Matthews, R James ^a   Dalton, Mark ^a   Roy, Garbiñe ^a   Chan, Andrew C ^a   Thomas, Matthew L ^a  

^a WASHINGTON UNIVERSITY SCHOOL OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

PROTEIN TYROSINE KINASE; PROTEIN TYROSINE PHOSPHATASE; T LYMPHOCYTE RECEPTOR;

ARTICLE; ENZYME ACTIVITY; PRIORITY JOURNAL; PROTEIN PHOSPHORYLATION; PROTEIN PROTEIN INTERACTION; T LYMPHOCYTE ACTIVATION;

EID: 0029904138     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.272.5265.1173     Document Type: Article

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9. 3, and 10 mM NaF]. Cell lysates for equal numbers of cells were separated on 8% SDS-potyacrylamide gel and immunoblotted with anti-phosphotyrosine (antibody 4G10, UBI). J.SHP-1 C/S 213 and J.neo are representative clones. 3L2 cells overexpressing SHP-1 or SHP-1 (C453S), or vector control cell lines, were generated by transfection of cells with either cytomegalovirus-driven expression vector (BCMG) containing SHP-1 or SHP-1 (C453S) cDNA, or vector with no insert. Neomycin-resistant cell lines were obtained, then individual clones were obtained by limiting dilution. Three individual clones were examined for each line. Equivalent expression of CDS on the resulting clones was determined by flow cytometric analysis. 3L2 hybridoma cells were stimulated for 24 hours with the hemoglobin β peptide(64-76) with CH27 cells used as antigen-presenting cells.
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27. We thank P. Allen and A. Shaw for advice and the gifts of reagents, and L. Dustin, M. Tsotsiashvili, and T. Woodford-Thomas for reading the manuscript. A.C C. is supported in part by the Arthritis Foundation and the Pews Scholars Program. D.R.P. is funded by the Lucille P. Markey Program. A.C.C. and M.LT. are investigators at the Howard Hughes Medical Institute.