title: Bacillolysin MA, a novel bacterial metalloproteinase that produces angiostatin-like fragments from plasminogen and activates protease zymogens in the coagulation and fibrinolysis systems.
abstract: We isolated a novel protease that converts plasminogen to angiostatin-like fragments (BL-angiostatins) from a culture of Bacillus megaterium A9542 through a single-step chromatography on CM-cellulose. The protease, designated bacillolysin MA (BL-MA), belongs to a family of neutral metalloproteinases based on the nucleotide sequence of its gene. At an enzyme:substrate ratio of 1:540, BL-MA cleaved human plasminogen mainly at Ser441-Val442 to form BL-angiostatin and miniplasminogen with a K(m) of 3.0 +/- 0.8 microM and a k(cat) of 0.70 +/- 0.09 s(-1). The resulting BL-angiostatins inhibited the proliferation, migration, and tube formation of vascular endothelial cells at concentrations of 1-10 microg/ml. Although BL-MA failed to activate plasminogen, it increased urokinase-catalyzed activation of plasminogen caused by production of miniplasminogen, which is highly susceptible to activation. In addition, BL-MA was active in converting prourokinase, prothrombin, coagulation factor X, and protein C to their active forms. BL-MA enhanced both the clotting of human plasma and clot dissolution in the presence of prourokinase. Thus, BL-MA affects blood coagulation and fibrinolysis systems and can be used to produce angiostatin-like plasminogen fragments and active serine proteases of human plasma.
authors.affiliation: Department of Applied Biological Science, Tokyo Noko University, 3-5-8 Saiwaicho, Fuchu-shi, Tokyo 183-8509, Japan.
authors.forename: Ritsuko
authors.initials: R
authors.lastname: Narasaki
authors.forename: Harushige
authors.initials: H
authors.lastname: Kuribayashi
authors.forename: Kosuke
authors.initials: K
authors.lastname: Shimizu
authors.forename: Daisuke
authors.initials: D
authors.lastname: Imamura
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Sato
authors.forename: Keiji
authors.initials: K
authors.lastname: Hasumi
mesh_terms: D000595:Amino Acid Sequence; D043168:Angiostatins / Q000378:metabolism*; D000818:Animals; D001410:Bacillus megaterium / Q000201:enzymology; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001777:Blood Coagulation / Q000502:physiology*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism*; D005164:Factor IX / Q000378:metabolism; D005170:Factor X / Q000378:metabolism; D005342:Fibrinolysis / Q000502:physiology*; D006801:Humans; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism*; D010958:Plasminogen / Q000235:genetics / Q000378:metabolism*; D011486:Protein C / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism; D011516:Prothrombin / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004792:Enzyme Precursors; D007527:Isoenzymes; D010446:Peptide Fragments; D011486:Protein C; D011498:Protein Precursors; D043168:Angiostatins; D011516:Prothrombin; D005164:Factor IX; D005170:Factor X; D010958:Plasminogen; D014568:Urokinase-Type Plasminogen Activator; D008666:Metalloendopeptidases; C071298:bacillolysin