The C-terminal domain promotes the hemorrhagic damage caused by Vibrio vulnificus metalloprotease.
Vibrio vulnificus, an opportunistic human pathogen, produces a 45-kDa zinc metalloprotease (V. vulnificus protease; VVP) as an important virulence determinant. VVP injected intradermally into the dorsal skin causes the hemorrhagic damage through specific degradation of type IV collage in the vascular basement membrane. The N-terminal 35-kDa polypeptide (VVP-N), the catalytic domain, also evoked the hemorrhagic skin reaction within minutes. However, the hemorrhagic activity of VVP-N was one-third of that of VVP. Besides, the proteolytic activity of VVP-N toward the reconstituted basement membrane or type IV collagen was found to be about 50 % of VVP. VVP-N, like VVP, was quickly inactivated by an equimolar amount of alpha(2)-macroglobulin, a broad-spectrum plasma protease inhibitor. These findings indicate that the C-terminal 10-kDa polypeptide, the substrate-binding domain mediating the effective binding to protein substrates, functions to augment the hemorrhagic reaction of VVP.
Miyoshi
S
S
Faculty of Pharmaceutical Sciences, Okayama University, Tsushima-Naka, Okayama 700-8530, Japan. miyoshi@pharm.okayama-u.ac.jp
Kawata
K
K
Tomochika
K
K
Shinoda
S
S
Yamamoto
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins; D001427:Bacterial Toxins / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D001485:Basement Membrane / Q000187:drug effects / Q000378:metabolism; D024141:Collagen Type IV / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006168:Guinea Pigs; D006470:Hemorrhage / Q000139:chemically induced* / Q000473:pathology; D007271:Injections, Intradermal; D008297:Male; D008666:Metalloendopeptidases / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins; D017386:Sequence Homology, Amino Acid; D012871:Skin Diseases / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology; D014733:Vibrio / Q000201:enzymology*; D000511:alpha-Macroglobulins / Q000494:pharmacology
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D024141:Collagen Type IV; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D000511:alpha-Macroglobulins; D008666:Metalloendopeptidases; C070101:vvP protein, Vibrio vulnificus