Purification and characterization of a vulnificolysin-like cytolysin produced by Vibrio tubiashii.
An extracellular cytolysin from Vibrio tubiashii was purified by sequential hydrophobic interaction chromatography with phenyl-Sepharose CL-4B and gel filtration with Sephacryl S-200. This protein is sensitive to heat and proteases, is inhibited by cholesterol, and has a molecular weight of 59,000 and an isoelectric point of 5.3. In addition to lysing various erythrocytes, it is cytolytic and/or cytotoxic to Chinese hamster ovary cells, Caco-2 cells, and Atlantic menhaden liver cells in tissue culture. Lysis of erythrocytes occurs by a multihit process that is dependent on temperature and pH. Twelve of the first 17 N-terminal amino acid residues (Asp-Asp-Tyr-Val-Pro-Val-Val-Glu-Lys-Val-Tyr-Tyr-Ile-Thr-Ser-Ser-Lys) are identical to those of the Vibrio vulnificus cytolysin.
Kothary
M H
MH
Divisions of Virulence Assessment, Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration, Washington, DC 20204, USA. mkothary@cfsan.fda.gov
Delston
R B
RB
Curtis
S K
SK
McCardell
B A
BA
Tall
B D
BD
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells / Q000187:drug effects; D018938:Caco-2 Cells / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects; D006224:Cricetinae; D003603:Cytotoxins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification* / Q000633:toxicity*; D004912:Erythrocytes / Q000187:drug effects; D005399:Fishes; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D008969:Molecular Sequence Data; D014733:Vibrio / Q000378:metabolism*
D003603:Cytotoxins