title: 1.2 A crystal structure of the serine carboxyl proteinase pro-kumamolisin; structure of an intact pro-subtilase.
abstract: Kumamolisin, an extracellular proteinase derived from an acido/thermophilic Bacillus, belongs to the sedolisin family of endopeptidases characterized by a subtilisin-like fold and a Ser-Glu-Asp catalytic triad. In kumamolisin, the Asp82 carboxylate hydrogen bonds to Glu32-Trp129, which might act as a proton sink stabilizing the catalytic residues. The 1.2/1.3 A crystal structures of the Glu32-->Ala and Trp129-->Ala mutants show that both mutations affect the active-site conformation, causing a 95% activity decrease. In addition, the 1.2 A crystal structure of the Ser278-->Ala mutant of pro-kumamolisin was determined. The prodomain exhibits a half-beta sandwich core docking to the catalytic domain similarly as the equivalent subtilisin prodomains in their catalytic-domain complexes. This pro-kumamolisin structure displays, for the first time, the uncleaved linker segment running across the active site and connecting the prodomain with the properly folded catalytic domain. The structure strongly points to an initial intramolecular activation cleavage in subtilases, as presumed for pro-subtilisin and pro-furin.
authors.affiliation: Department of Structure Research, Max-Planck-Institute for Biochemistry, Am Klopferspitz 18a, D-82152 Martinsried, Germany.
authors.forename: Mireia
authors.initials: M
authors.lastname: Comellas-Bigler
authors.forename: Klaus
authors.initials: K
authors.lastname: Maskos
authors.forename: Robert
authors.initials: R
authors.lastname: Huber
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Oyama
authors.forename: Kohei
authors.initials: K
authors.lastname: Oda
authors.forename: Wolfram
authors.initials: W
authors.lastname: Bode
mesh_terms: D000595:Amino Acid Sequence; D016282:Aspartic Acid Endopeptidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology*; D001665:Binding Sites; D018360:Crystallography, X-Ray; D003625:Data Collection; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017510:Protein Folding; D040681:Structural Homology, Protein; D020860:Subtilisin / Q000737:chemistry / Q000378:metabolism
chemical_list: D020860:Subtilisin; D016282:Aspartic Acid Endopeptidases; C078894:kumamolysin