title: Secreted neutral metalloproteases of Bacillus anthracis as candidate pathogenic factors.
abstract: To evaluate the pathogenic potential of Bacillus anthracis-secreted proteases distinct from lethal toxin, two neutral zinc metalloproteases were purified to apparent homogeneity from the culture supernatant of a non-virulent delta Ames strain (pXO1-, pXO2-). The first (designated Npr599) is a thermolysin-like enzyme highly homologous to bacillolysins from other Bacillus species. The second (designated InhA) is a homolog of the Bacillus thuringiensis immune inhibitor A. These proteases belong to the M4 and M6 families, respectively. Both enzymes digested various substrates, including extracellular matrix proteins, endogenous inhibitors, and coagulation proteins, with some differences in specificity. In addition, InhA accelerated urokinase-mediated plasminogen activation, suggesting that InhA acts as a modulator of plasmin in the host inflammatory system. Relevant to epithelial barrier function, Npr599 and InhA significantly enhanced syndecan-1 shedding from cultured normal murine mammary gland cells without affecting their viability through stimulation of the host cell ectodomain shedding mechanism. In addition, Npr599 and InhA directly cleaved recombinant syndecan-1 fused to glutathione S-transferase. Mass spectrometric analysis suggested that the cleavage sites of Npr599 and InhA are the Asp(39)-Asp(40) and Gly(48)-Thr(49) bonds, respectively. We propose that Npr599 and InhA from B. anthracis are multifunctional pathogenic factors that may contribute to anthrax pathology through direct degradation of host tissues, increases in barrier permeability, and/or modulation of host defenses.
authors.affiliation: National Center for Biodefense and Infectious Diseases, George Mason University, Manassas, Virginia 20110, USA.
authors.forename: Myung-Chul
authors.initials: MC
authors.lastname: Chung
authors.forename: Taissia G
authors.initials: TG
authors.lastname: Popova
authors.forename: Bryan A
authors.initials: BA
authors.lastname: Millis
authors.forename: Dhritiman V
authors.initials: DV
authors.lastname: Mukherjee
authors.forename: Weidong
authors.initials: W
authors.lastname: Zhou
authors.forename: Lance A
authors.initials: LA
authors.lastname: Liotta
authors.forename: Emanuel F
authors.initials: EF
authors.lastname: Petricoin
authors.forename: Vikas
authors.initials: V
authors.lastname: Chandhoke
authors.forename: Charles
authors.initials: C
authors.lastname: Bailey
authors.forename: Serguei G
authors.initials: SG
authors.lastname: Popov
mesh_terms: D000818:Animals; D001408:Bacillus anthracis / Q000201:enzymology* / Q000472:pathogenicity*; D002478:Cells, Cultured; D017364:Collagenases / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D015964:Gene Expression Regulation, Bacterial; D005982:Glutathione Transferase / Q000378:metabolism; D007249:Inflammation; D007700:Kinetics; D013058:Mass Spectrometry; D045726:Metalloproteases / Q000737:chemistry*; D051379:Mice; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D011994:Recombinant Proteins; D005982:Glutathione Transferase; D045726:Metalloproteases; D017364:Collagenases