title: Association of CD45 with Lck and components of the Ras signalling pathway in pervanadate-treated mouse T-cell lines.
abstract: Pervanadate treatment of a mouse T-cell hybridoma cell line overexpressing an activated form of p56lck was shown to result in tyrosine phosphorylation of CD45. Immunoprecipitates prepared under mild lysis conditions using antibodies against CD45 contained a number of other proteins, including p56lck, that were not evident in the absence of pervanadate treatment or in T cells lacking activated Lck, implying that under these conditions, CD45 is present within complexes containing Lck and other proteins. Analyses involving deletion mutants of p56lck indicated that interactions with CD45 did not absolutely require the SH2 and SH3 regions of Lck. Three proteins of the Ras signalling pathway were also shown to associated with CD45: the GTPase-activating protein for Ras (rasGAP), the signalling protein Grb2, and, possibly via complex formation with Grb2, the guanine nucleotide exchange factor mammalian son of sevenless (mSOS). In addition, CD45 was also found in immunoprecipitates prepared from these cells using an antiserum which recognizes Vav. It is possible that rasGAP, Grb2 and Vav bind to phosphotyrosine residues on CD45 via SH2 domains, and such interactions may be specific as other SH2-containing proteins, including phospholipase C alpha (PLC gamma), the p85 subunit of phosphatidylinositol 3-kinase (PI 3-kinase). She and Syp/PTP1D were not detectably associated with CD45 under the same conditions. These data suggested that in addition to its role as a protein tyrosine phosphatase, CD45 may participate in T-cell activation by serving as a membrane docking site for components of the Ras signalling pathway.
authors.affiliation: Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
authors.forename: J M
authors.initials: JM
authors.lastname: Lee
authors.forename: M
authors.initials: M
authors.lastname: Fournel
authors.forename: A
authors.initials: A
authors.lastname: Veillette
authors.forename: P E
authors.initials: PE
authors.lastname: Branton
mesh_terms: D000818:Animals; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D017493:Leukocyte Common Antigens / Q000502:physiology*; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D051379:Mice; D010766:Phosphorylation; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors*; D016283:Proto-Oncogene Proteins p21(ras) / Q000502:physiology*; D051381:Rats; D013601:T-Lymphocytes / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism; D014638:Vanadates / Q000494:pharmacology*; D018909:src Homology Domains; D019061:src-Family Kinases / Q000502:physiology*
chemical_list: D004791:Enzyme Inhibitors; C076715:pervanadate; D014638:Vanadates; D014443:Tyrosine; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D019061:src-Family Kinases; D017493:Leukocyte Common Antigens; D017027:Protein Tyrosine Phosphatases; D016283:Proto-Oncogene Proteins p21(ras)