title: The structure of jack bean urease. The complete amino acid sequence, limited proteolysis and reactive cysteine residues.
abstract: The amino acid sequence of jack bean urease has been determined. The protein consists of a single kind of polypeptide chain containing 840 amino acid residues. The subunit relative molecular mass calculated from the sequence is 90,770, indicating that urease is composed of six subunits. Out of 25 histidine residues in urease, 13 were crowded in the region between residues 479 and 607, suggesting that this region may contain the nickel-binding site. Limited tryptic digestion cleaved urease at two sites, Lys-128 and Lys-662. Proteolytic products were not dissociated and retained full enzymatic activity. Five tryptic peptides containing the reactive cysteine residues were isolated and characterized with the aid of sulfhydryl-specific reagents, N-iodoacetyl-N'-(5-sulfo-1-naphthyl)ethylenediamine and N-(7-dimethylamino-4-methyl-3-coumarinyl)-maleimide. The reactive cysteine residues were located at positions 59, 207, 592, 663, and 824. The possibility that Cys-59, Cys-207, Cys-663, and Cys-824 are involved in the urease activity of the enzyme has been eliminated. Cys-592, which is essential for enzymatic activity, is located in the above-mentioned histidine-rich region.
authors.affiliation: Department of Biochemistry, National Defense Medical College, Tokorozawa, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Takishima
authors.forename: T
authors.initials: T
authors.lastname: Suga
authors.forename: G
authors.initials: G
authors.lastname: Mamiya
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D003545:Cysteine / Q000032:analysis*; D007887:Fabaceae / Q000201:enzymology; D008969:Molecular Sequence Data; D009532:Nickel / Q000378:metabolism; D010449:Peptide Mapping; D010944:Plants / Q000201:enzymology*; D010946:Plants, Medicinal; D014510:Urease / Q000032:analysis*
chemical_list: D009532:Nickel; D014510:Urease; D003545:Cysteine