Biological properties of the venom from the scorpionfish (Scorpaena plumieri) and purification of a gelatinolytic protease.
In this work we describe some biological properties and a partial biochemical characterization of the Scorpanea plumieri crude venom. The fresh venom induced a decrease in blood pressure, cardiac and respiratory frequency, and exhibited hemorrhagic, hemolytic and proteolytic activities. The LD(50) (i.v. mouse) was 0.28 mg/kg. The pharmacological activities were found to be very unstable and this fact could be associated with proteolytic activity. Enzymes which hydrolyze casein and gelatin were found in this venom. A gelatinolytic protease (Sp-GP) was purified to homogeneity from S. plumieri venom through a combination of three chromatographic steps: gel filtration on Sephacryl S-200; ion exchange on DEAE-cellulose and reverse-phase/HPLC on a Vydac C4 column. The purified protease was approximately 2% of the whole protein in the soluble crude venom. The molecular mass of the Sp-GP scorpionfish gelatinase estimated by SDS-PAGE was around 80,000 Da under reducing conditions and 72,000 Da under non-reducing conditions. Attempts to determine the N-terminal sequence by automatic Edman degradation were unsuccessful, probably due to blockage of the N-terminal group. Gelatinolytic activity was optimal at pH 7-8. This is the first report of the isolation and characterization of a scorpionfish venom protease.
Carrijo
Linda Christian
LC
Departamento de Ciências Fisiológicas, Centro Biomédico, Universidade Federal do Espírito Santo, Vitória, ES, Brazil.
Andrich
Filipe
F
de Lima
Maria Elena
ME
Cordeiro
Marta N
MN
Richardson
Michael
M
Figueiredo
Suely G
SG
D000818:Animals; D002364:Caseins / Q000187:drug effects; D004912:Erythrocytes / Q000187:drug effects / Q000502:physiology; D005397:Fish Venoms / Q000201:enzymology* / Q000302:isolation & purification / Q000494:pharmacology*; D005400:Fishes, Poisonous; D005780:Gelatin / Q000187:drug effects*; D006461:Hemolysis; D006470:Hemorrhage / Q000139:chemically induced; D007022:Hypotension / Q000139:chemically induced; D007928:Lethal Dose 50; D008297:Male; D051379:Mice; D010447:Peptide Hydrolases / Q000302:isolation & purification*; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D012867:Skin / Q000187:drug effects / Q000473:pathology
D002364:Caseins; D005397:Fish Venoms; C002087:N,N-dimethylcasein; D005780:Gelatin; D010447:Peptide Hydrolases