title: A single cysteine, Cys-64, is essential for assembly of tenascin-C hexabrachions.
abstract: Tenascin-C is a large, multimeric extracellular matrix protein that is found in a variety of tissues and can have profound effects on cell adhesion. It is secreted from cells as a hexamer of six identical chains called a hexabrachion. Disulfide bonding among tenascin subunits mediates intracellular assembly into hexamers. The amino-terminal assembly domain consists of heptad repeats and at least six cysteine residues (Cys-64, -111, -113, -140, -146, -147) that could be involved in multimerization. We have now determined the requirements for these cysteine residues during hexamer assembly. Our results show that only Cys-64 is required to form the hexameric structure. Mutation of Cys-64 to glycine resulted in release of trimer intermediates, which probably form via the heptad repeats, but no hexamers were secreted. In contrast, individual or pairs of mutations of each of the other cysteines had no effect on tenascin hexamer formation, and inclusion of any other cysteine mutations along with C64G did not further disrupt the multimer pattern. However, when all six cysteines were mutated, monomers were the major extracellular form. Together, these results show that trimers are an intermediate of tenascin-C assembly and that Cys-64 is essential for formation of hexabrachions.
authors.affiliation: Department of Molecular Biology, Princeton University, New Jersey 08544-1014, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Luczak
authors.forename: S D
authors.initials: SD
authors.lastname: Redick
authors.forename: J E
authors.initials: JE
authors.lastname: Schwarzbauer
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D019556:COS Cells; D002478:Cells, Cultured; D003545:Cysteine / Q000235:genetics / Q000378:metabolism*; D004220:Disulfides / Q000032:analysis; D051379:Mice; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D019063:Tenascin / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D004220:Disulfides; D019063:Tenascin; D003545:Cysteine