title: Proteolytic fragmentation of tetanus toxin by subcellular fractions of JY, a B lymphoblastoid cell line.
abstract: Proteolysis of 125I-labelled tetanus toxin by subcellular fractions from an Epstein-Barr-virus-transformed B lymphoblastoid cell line, JY, was investigated. Fractions enriched in lysosomes and plasma membranes cleaved the toxin molecule at several sites, with a pH optimum of 5.5. N-Terminal sequence analysis of Mr-81,000, -45,000 and -35,000 proteolytic fragments indicated cleavage of the Asp-460-Leu-461, Asp-872-Glu-873 and Ile-1013-Thr-1014 peptide bonds, all sites located within the heavy chain of the toxin molecule. Additional sites near the C-terminus of the heavy chain, giving rise to low-Mr peptides, were implicated. The toxin light chain was more resistant to proteolysis. A similar pattern of fragmentation was observed with tetanus toxin biosynthetically radiolabelled with 14C-labelled amino acids, showing that the proteolysis was not an artifact caused by iodination. The proteolytic activity was inhibited by the serine proteinase inhibitor di-isopropyl phosphorofluoridate, thiol-blocking proteinase inhibitors N-ethylmaleimide and iodoacetamide, and by EDTA. These results represent a preliminary characterization of the processing in vitro of tetanus toxin by an antigen-presenting cell line.
authors.affiliation: DBMS/ICH, Unité INSERM 238, CEN-G BP 85X, Grenoble, France.
authors.forename: A
authors.initials: A
authors.lastname: Reboul
authors.forename: J
authors.initials: J
authors.lastname: Arvieux
authors.forename: J F
authors.initials: JF
authors.lastname: Wright
authors.forename: M G
authors.initials: MG
authors.lastname: Colomb
mesh_terms: D001402:B-Lymphocytes; D002458:Cell Fractionation; D002460:Cell Line; D002472:Cell Transformation, Viral; D004591:Electrophoresis, Polyacrylamide Gel; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D010446:Peptide Fragments / Q000302:isolation & purification; D013347:Subcellular Fractions / Q000378:metabolism; D013744:Tetanus Toxin / Q000378:metabolism*
chemical_list: D046911:Macromolecular Substances; D010446:Peptide Fragments; D013744:Tetanus Toxin