title: Proteolytic fragmentation of sialophorin (CD43). Localization of the activation-inducing site and examination of the role of sialic acid.
abstract: Sialophorin (CD43) is the major surface mucin on many hematopoietic cells. It has been implicated in regulating the survival of T lymphocytes in the circulation, and its functions in vitro as the receptor of a T lymphocyte and monocyte activation pathway. The structure of CD43 was examined by protease treatment of lymphoblastoid cells bearing surface CD43. Trypsin treatment converts CD43 (apparent Mr 115,000) to species of apparent Mr 100,000 called T-100, which remains cell-associated; however, the mechanism of trypsin action was not clarified. Pancreatic elastase and Staphylococcus aureus V8 protease cleave CD43 at discrete extracellular sites. V8 protease generates two fragments, which together account for all properties and mass of the parent molecule. The COOH-terminal fragment V-90 (apparent Mr 90,000) consists of the intracellular and transmembrane regions and part of the extracellular region. The fragment V-30 (apparent Mr 30,000), which is released from the cell, comprises the NH2-terminal approximately 78 amino acids with attached oligosaccharides. V-30 contains the binding sites for the antibodies L2 and L10; the latter is the antibody that activates lymphocytes and monocytes. These findings subdivide the extracellular region of CD43 and indicate that the activation-inducing epitope is located in the most distal portion of the molecule. It is shown that CD43 is insensitive to all but very high concentrations of three proteases. Pretreatment with sialidase enhances sensitivity 13-fold for trypsin, 40-fold for S. aureus V8 protease, and 400-fold for elastase, suggesting that sialic acid influences the survival of surface CD43 molecules when cells are exposed to protease.
authors.affiliation: Center for Blood Research, Children's Hospital, Boston, MA 02115.
authors.forename: E
authors.initials: E
authors.lastname: Remold-O'Donnell
authors.forename: F S
authors.initials: FS
authors.lastname: Rosen
mesh_terms: D015703:Antigens, CD; D002478:Cells, Cultured; D006801:Humans; D051917:Leukosialin; D008213:Lymphocyte Activation; D009000:Monocytes / Q000276:immunology; D019158:N-Acetylneuraminic Acid; D010196:Pancreatic Elastase / Q000494:pharmacology; D010446:Peptide Fragments / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000494:pharmacology; D012794:Sialic Acids / Q000502:physiology*; D012795:Sialoglycoproteins / Q000378:metabolism*; D014357:Trypsin / Q000494:pharmacology
chemical_list: D015703:Antigens, CD; D051917:Leukosialin; D010446:Peptide Fragments; C465014:SPN protein, human; D012794:Sialic Acids; D012795:Sialoglycoproteins; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D010196:Pancreatic Elastase; D014357:Trypsin; D019158:N-Acetylneuraminic Acid