Evidence that leukosialin, CD43, is intensely sulfated in the murine T lymphoma line RDM-4.
Metabolic labeling of the murine T lymphoma cell line RDM-4 with [35S] sulfate results in intense incorporation into a cell-retained component of apparent Mr approximately 100,000. This macromolecule is identified as a glycoprotein by lectin chromatography. The sulfate is not incorporated as tyrosine sulfate. Release of the radiolabel by alkaline beta-elimination but not by endoglycosidase F is consistent with the sulfation of O- rather than N-linked oligosaccharides. The sulfated glycoprotein displays anomalous migration on SDS-PAGE in two respects: 1) the apparent Mr shifts from 115,000 to 87,000 on increasing the acrylamide concentration from 7 to 12%, and 2) on neuraminidase digestion migration is substantially reduced (apparent Mr 140,000). These properties indicate that the sulfated protein is both heavily glycosylated and extensively sialylated, and are characteristic of the lymphoid mucin, leukosialin (sialophorin, CD43). Specific labeling of the sialoglycoproteins of RDM-4 cells indicates that leukosialin, the most intensely labeled protein, comigrates with the sulfated protein on SDS-PAGE at varying acrylamide concentrations. Our data are therefore consistent with sulfation of at least some of the numerous O-linked oligosaccharides of this abundant glycoprotein in RDM-4 cells. No sulfation of CD43 in resting splenic T cells is observed.
Wilson
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AP
Department of Biochemistry, Royal Holloway and Bedford New College, University of London, Egham, Surrey, UK.
Rider
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CC
D000818:Animals; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte / Q000737:chemistry; D066298:In Vitro Techniques; D051917:Leukosialin; D051379:Mice; D008970:Molecular Weight; D009439:Neuraminidase / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D012795:Sialoglycoproteins / Q000737:chemistry*; D013431:Sulfates / Q000378:metabolism*; D013601:T-Lymphocytes / Q000737:chemistry*; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D051917:Leukosialin; D012795:Sialoglycoproteins; C497863:Spn protein, mouse; D013431:Sulfates; D009439:Neuraminidase