title: Two thimet oligopeptidase-like Pz peptidases produced by a collagen-degrading thermophile, Geobacillus collagenovorans MO-1.
abstract: A collagen-degrading thermophile, Geobacillus collagenovorans MO-1, was found to produce two metallopeptidases that hydrolyze the synthetic substrate 4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg (Pz-PLGPR), containing the collagen-specific sequence -Gly-Pro-X-. The peptidases, named Pz peptidases A and B, were purified to homogeneity and confirmed to hydrolyze collagen-derived oligopeptides but not collagen itself, indicating that Pz peptidases A and B contribute to collagen degradation in collaboration with a collagenolytic protease in G. collagenovorans MO-1. There were many similarities between Pz peptidases A and B in their catalytic properties; however, they had different molecular masses and shared no antigenic groups against the respective antibodies. Their primary structures clarified from the cloned genes showed lower identity (22%). From homology analysis for proteolytic enzymes in the database, the two Pz peptidases belong to the M3B family. In addition, Pz peptidases A and B shared high identities of over 70% with unassigned peptidases and oligopeptidase F-like peptidases of the M3B family, respectively. Those homologue proteins are putative in the genome database but form two distinct segments, including Pz peptidases A and B, in the phylogenic tree. Mammalian thimet oligopeptidases, which were previously thought to participate in collagen degradation and share catalytic identities with Pz peptidases, were found to have lower identities in the overall primary sequence with Pz peptidases A and B but a significant resemblance in the vicinity of the catalytic site.
authors.affiliation: Department of Applied Biochemistry, Kyoto Prefectural University, Shimogamo, Sakyo, Kyoto 606-8522, Japan. .
authors.forename: Ryoma
authors.initials: R
authors.lastname: Miyake
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Shigeri
authors.forename: Yoshiro
authors.initials: Y
authors.lastname: Tatsu
authors.forename: Noboru
authors.initials: N
authors.lastname: Yumoto
authors.forename: Midori
authors.initials: M
authors.lastname: Umekawa
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Tsujimoto
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Matsui
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Watanabe
mesh_terms: D000595:Amino Acid Sequence; D001419:Bacteria / Q000201:enzymology*; D001426:Bacterial Proteins / Q000378:metabolism; D001483:Base Sequence; D002876:Chromosomes, Bacterial; D003094:Collagen / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008666:Metalloendopeptidases / Q000096:biosynthesis* / Q000737:chemistry*; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D003094:Collagen; D008666:Metalloendopeptidases; C022510:thimet oligopeptidase