BjussuSP-I: a new thrombin-like enzyme isolated from Bothrops jararacussu snake venom.
A thrombin-like enzyme named BjussuSP-I, isolated from B. jararacussu snake venom, is an acidic single chain glycoprotein with approximately 6% sugar, Mr=61,000 under reducing conditions and pI approximately 3.8, representing 1.09% of the chromatographic A(280) recovery. BjussuSP-I is a glycosylated serine protease containing both N-linked carbohydrates and sialic acid in its structure. BjussuSP-I showed a high clotting activity upon human plasma, which was inhibited by PMSF, leupeptin, heparin and 1,10-phenantroline. This enzyme showed high stability regarding coagulant activity when analyzed at different temperatures (-70 to 37 degrees C), pHs (4.5 to 8.0), and presence of two divalent metal ions (Ca(2+) and Mg(2+)). It also displayed TAME esterase and proteolytic activities toward natural (fibrinogen and fibrin) and synthetic (BAPNA) substrates, respectively, being also inhibited by PMSF and leupeptin. BjussuSP-I can induce production of polyclonal antibodies able to inhibit its clotting activity, but unable to inhibit its proteolytic activity on fibrinogen. The enzyme also showed crossed immunoreactivity against 11 venom samples of Bothrops, 1 of Crotalus, and 1 of Calloselasma snakes, in addition of LAAO isolated from B. moojeni venom. It displayed neither hemorrhagic, myotoxic, edema-inducing profiles nor proteolytic activity on casein. BjussuSP-I showed an N-terminal sequence (VLGGDECDINEHPFLA FLYS) similar to other thrombin-like enzymes from snake venoms. Based on its biochemical, enzymatic and pharmacological characteristics, BjussuSP-I was identified as a new thrombin-like enzyme isoform from Bothrops jararacussu snake venom.
Sant' Ana
Carolina D
CD
Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Universidade de São Paulo, FCFRP-USP, Ribeirão Preto - SP, Brazil.
Ticli
Fabio K
FK
Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Universidade de São Paulo, FCFRP-USP, Ribeirão Preto - SP, Brazil.
Oliveira
Leandro L
LL
Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto - Universidade de São Paulo, FMRP-USP, Ribeirão Preto - SP, Brazil.
Giglio
Jose R
JR
Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto - Universidade de São Paulo, FMRP-USP, Ribeirão Preto - SP, Brazil.
Rechia
Carem G V
CGV
Departamento de Física e Química, Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Universidade de São Paulo, FCFRP-USP, Ribeirão Preto - SP, Brazil.
Fuly
André L
AL
Instituto de Biologia, Departamento de Biologia Celular e Molecular, Universidade Federal Fluminense, UFF, Niterói, RJ, Brazil.
Selistre de Araújo
Heloisa S
HS
Departamento de Ciências Fisiológicas, Universidade Federal de São Carlos, UFSCar, São Carlos, SP, Brazil.
Franco
João J
JJ
Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Universidade de São Paulo, FCFRP-USP, Ribeirão Preto - SP, Brazil.
Stabeli
Rodrigo G
RG
Laboratório de Bioquímica e Biotecnologia, Instituto de Pesquisas em Patologias Tropicais (IPEPATRO), Universidade de Rondônia, UNIR, Porto Velho-RO, Brazil.
Soares
Andreimar M
AM
Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Universidade de São Paulo, FCFRP-USP, Ribeirão Preto - SP, Brazil. Electronic address: andreims@fcfrp.usp.br.
Sampaio
Suely V
SV
Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Universidade de São Paulo, FCFRP-USP, Ribeirão Preto - SP, Brazil. Electronic address: suvilela@fcfrp.usp.br.
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001777:Blood Coagulation; D017837:Bothrops / Q000378:metabolism*; D002845:Chromatography; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology* / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D011817:Rabbits; D012697:Serine Endopeptidases / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D013917:Thrombin / Q000737:chemistry*
D000906:Antibodies; D003435:Crotalid Venoms; D012697:Serine Endopeptidases; D013917:Thrombin