Molecular characterization of BjussuSP-I, a new thrombin-like enzyme with procoagulant and kallikrein-like activity isolated from Bothrops jararacussu snake venom.
A thrombin-like enzyme, named BjussuSP-I, isolated from Bothrops jararacussu snake venom, is an acidic single-chain glycoprotein with M(r)=61,000, pI approximately 3.8 and 6% sugar. BjussuSP-I shows high proteolytic activity upon synthetic substrates, such as S-2238 and S-2288. It also shows procoagulant and kallikrein-like activity, but is unable to act on platelets and plasmin. These activities are inhibited by specific inhibitors of this class of enzymes. The complete cDNA sequence of BjussuSP-I with 696bp encodes open reading frames of 232 amino acid residues, which conserve the common domains of thrombin-like serine proteases. BjussuSP-I shows a high structural homology with other thrombin-like enzymes from snake venoms where common amino acid residues are identified as those corresponding to the catalytic site and subsites S1, S2 and S3 already reported. In this study, we also demonstrated the importance of N-linked glycans to improve thrombin-like activity of BjussuSP-I toxin.
Sant'Ana
Carolina D
CD
Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto da Universidade de São Paulo, FCFRP-USP, Ribeirão Preto, SP, Brazil.
Bernardes
Carolina P
CP
Izidoro
Luiz Fernando M
LF
Mazzi
Maurício V
MV
Soares
Sandro G
SG
Fuly
André L
AL
Zingali
Russolina B
RB
Magro
Angelo J
AJ
Braz
Antonio S K
AS
Fontes
Marcos R M
MR
Stábeli
Rodrigo G
RG
Sampaio
Suely V
SV
Soares
Andreimar M
AM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001779:Blood Coagulation Factors / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D017837:Bothrops; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000201:enzymology*; D018076:DNA, Complementary / Q000378:metabolism; D007610:Kallikreins / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D013917:Thrombin / Q000737:chemistry; D013918:Thrombin Time
D001779:Blood Coagulation Factors; D003435:Crotalid Venoms; D018076:DNA, Complementary; D007610:Kallikreins; D012697:Serine Endopeptidases; D013917:Thrombin