Characterization of lineage restricted forms of a Xenopus CD45 homologue.
The leukocyte common antigen, also known as CD45, is a structurally heterogenous molecule ranging in molecular weight from 180 to 220 kDa. CD45 belongs to a family of high molecular weight, cell surface glycoproteins expressed on all hematopoietic lineages with the exception of mature erythrocytes. In higher vertebrates, the highly conserved cytoplasmic domain of CD45 exhibits protein tyrosine phosphatase activity and has been implicated in lymphocyte activation through dephosphorylation of critical tyrosine residues on substrates associated with signal transduction pathways. The monoclonal antibody CL21 recognizes a high molecular weight determinant expressed on the surface of Xenopus leukocytes which was postulated to be a CD45 homologue. In order to determine if lymphocyte subpopulations expressed different molecular weight variants, splenic B cells were identified and isolated on the basis of surface IgM and the CL21 determinant expressed by these cells was compared to the determinant expressed by thymocytes. Immunoprecipitation revealed that IgM + B cells expressed a 220 kDa molecular weight variant whereas thymocytes and IgM-cells expressed a 180 kDa variant. Bone marrow myeloid cells, isolated on the basis of light scatter properties, expressed a determinant which ranged from 150 to 160 kDa. Dephosphorylation experiments utilizing p-nitrophenyl phosphate, 32P-labeled Raytide [tyr(P)], or Kemptide [ser(P)] as substrates demonstrated that immunoprecipitated CL21 antigen exhibited tyrosine specific phosphatase activity which was inhibited by sodium orthovanadate. Thus, data based on the presence of enzymatic activity and lineage restricted molecular weight variants support the hypothesis that the CL21 determinant is the amphibian homologue of mammalian CD45, and suggest that both structural and functional elements of CD45 have been conserved during vertebrate evolution.
Barritt
L C
LC
Department of Cell Biology and Anatomy, University of Nebraska Medical Center, Omaha 68198-6395, USA.
Turpen
J B
JB
D000818:Animals; D001402:B-Lymphocytes / Q000737:chemistry* / Q000201:enzymology / Q000276:immunology; D001853:Bone Marrow / Q000276:immunology; D001854:Bone Marrow Cells; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000201:enzymology / Q000276:immunology*; D017493:Leukocyte Common Antigens / Q000737:chemistry* / Q000502:physiology; D051379:Mice; D008970:Molecular Weight; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry / Q000276:immunology; D013154:Spleen / Q000737:chemistry / Q000166:cytology; D013379:Substrate Specificity; D013950:Thymus Gland / Q000737:chemistry; D014981:Xenopus / Q000276:immunology*
D017493:Leukocyte Common Antigens; D017027:Protein Tyrosine Phosphatases