Chemical modification of Vibrio vulnificus metalloprotease with activated polyethylene glycol.
Vibrio vulnificus, an opportunistic human pathogen causing septicemia, produces a metalloprotease which is suspected to be a virulence determinant, but which is labile in vivo due to inactivation by alpha-macroglobulin. To obtain a derivative which is stable in vivo, the metalloprotease was modified with activated monomethoxy polyethylene glycol. The modified protease retained full activity to a peptide substrate and 10-20% activity to protein substrates, and was resistant to entrapment by alpha-macroglobulin because of the increased molecular size (approx. 90 kDa). These findings suggest that the modified protease is stable in vivo and may be used to investigate the pathological actions of the protease in the bloodstream.
Narukawa
H
H
Faculty of Pharmaceutical Sciences, Okayama University, Japan.
Miyoshi
S
S
Shinoda
S
S
D016470:Bacteremia / Q000209:etiology; D004795:Enzyme Stability; D006801:Humans; D007120:Immunochemistry; D066298:In Vitro Techniques; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000276:immunology; D008970:Molecular Weight; D011092:Polyethylene Glycols; D014733:Vibrio / Q000201:enzymology* / Q000472:pathogenicity; D014735:Vibrio Infections / Q000209:etiology; D014774:Virulence; D000511:alpha-Macroglobulins / Q000494:pharmacology
D000511:alpha-Macroglobulins; D011092:Polyethylene Glycols; D008666:Metalloendopeptidases