Isolation and characterization of a serine proteinase with thrombin-like activity from the venom of the snake Bothrops asper.
A serine proteinase with thrombin-like activity was isolated from the venom of the Central American pit viper Bothrops asper. Isolation was performed by a combination of affinity chromatography on aminobenzamidine-Sepharose and ion-exchange chromatography on DEAE-Sepharose. The enzyme accounts for approximately 0.13% of the venom dry weight and has a molecular mass of 32 kDa as determined by SDS-PAGE, and of 27 kDa as determined by MALDI-TOF mass spectrometry. Its partial amino acid sequence shows high identity with snake venom serine proteinases and a complete identity with a cDNA clone previously sequenced from this species. The N-terminal sequence of the enzyme is VIGGDECNINEHRSLVVLFXSSGFL CAGTLVQDEWVLTAANCDSKNFQ. The enzyme induces clotting of plasma (minimum coagulant dose = 4.1 microg) and fibrinogen (minimum coagulant dose = 4.2 microg) in vitro, and promotes defibrin(ogen)ation in vivo (minimum defibrin(ogen)ating dose = 1.0 microg). In addition, when injected intravenously in mice at doses of 5 and 10 microg, it induces a series of behavioral changes, i.e., loss of the righting reflex, opisthotonus, and intermittent rotations over the long axis of the body, which closely resemble the ;gyroxin-like' effect induced by other thrombin-like enzymes from snake venoms.
Pérez
A V
AV
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Rucavado
A
A
Sanz
L
L
Calvete
J J
JJ
Gutiérrez
J M
JM
D000595:Amino Acid Sequence; D000818:Animals; D000997:Antivenins / Q000627:therapeutic use; D001777:Blood Coagulation / Q000187:drug effects; D017837:Bothrops; D002847:Chromatography, Agarose; D002852:Chromatography, Ion Exchange; D003029:Coagulants / Q000008:administration & dosage / Q000302:isolation & purification* / Q000494:pharmacology; D003364:Costa Rica; D003435:Crotalid Venoms / Q000201:enzymology*; D004353:Drug Evaluation, Preclinical; D005340:Fibrinogen / Q000378:metabolism; D051379:Mice; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D012909:Snake Bites / Q000503:physiopathology; D013917:Thrombin / Q000737:chemistry
D000997:Antivenins; D003029:Coagulants; D003435:Crotalid Venoms; D005340:Fibrinogen; D012697:Serine Endopeptidases; D013917:Thrombin