title: An intra-dimeric crosslink of large subunits of spinach ribulose-1,5-bisphosphate carboxylase/oxygenase is formed by oxidation of cysteine 247.
abstract: Crystals of the hexadecameric form of ribulose-bisphosphate carboxylase used to solve the structure of the enzyme are composed of protein substantially crosslinked by a disulfide bond between pairs of large subunits. Conditions leading to the selective formation of dimers of the large subunits are described. The stability and specificity of the intra-dimeric crosslink was used to confirm that only one cysteine residue, Cys247 of neighboring large subunits, is involved in the bridge. The ability to generate this disulfide selectively, or alternatively replace the cysteine by site-directed mutagenesis, has led us to conclude that there is no effect of these changes on any of the critical kinetic parameters of the enzyme. The benign effect of the oxidation indicates that the crystal structures of the ribulose-bisphosphate carboxylase, particularly of the active site, are a true representation of the native enzyme.
authors.affiliation: Central Research and Development Department, E.I. DuPont de Nemours and Company, Experimental Station Wilmington.
authors.forename: B
authors.initials: B
authors.lastname: Ranty
authors.forename: G
authors.initials: G
authors.lastname: Lorimer
authors.forename: S
authors.initials: S
authors.lastname: Gutteridge
mesh_terms: D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents; D003545:Cysteine / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010084:Oxidation-Reduction; D010449:Peptide Mapping; D010944:Plants / Q000201:enzymology; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000737:chemistry; D014357:Trypsin
chemical_list: D003432:Cross-Linking Reagents; D013438:Sulfhydryl Compounds; D014357:Trypsin; D012273:Ribulose-Bisphosphate Carboxylase; D003545:Cysteine