Molecular and functional characterization of a new non-hemorrhagic metalloprotease from Bothrops jararacussu snake venom with antiplatelet activity.
BjussuMP-II is an acidic low molecular weight metalloprotease (Mr approximately 24,000 and pI approximately 6.5), isolated from Bothrops jararacussu snake venom. The chromatographic profile in RP-HPLC and its N-terminal sequence confirmed its high purity level. Its complete cDNA was obtained by RT-PCR and the 615bp codified for a mature protein of 205 amino acid residues. The multiple alignment of its deduced amino acid sequence and those of other snake venom metalloproteases showed a high structural similarity, mainly among class P-I proteases. The molecular modeling analysis of BjussuMP-II showed also conserved structural features with other SVMPs. BjussuMP-II did not induce hemorrhage, myotoxicity and lethality, but displayed dose-dependent proteolytic activity on fibrinogen, collagen, fibrin, casein and gelatin, keeping stable at different pHs, temperatures and presence of several divalent ions. BjussuMP-II did not show any clotting or anticoagulant activity on human citrated plasma, in contrast to its inhibitory effects on platelet aggregation. The aspects broached, in this work, provide data on the relationship between structure and function, in order to better understand the effects elicited by snake venom metalloproteases.
Marcussi
Silvana
S
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
Santos-Filho
Norival A
NA
Mazzi
Maurício V
MV
Oliveira
Clayton Z
CZ
Izidoro
Luiz Fernando M
LF
Fuly
André L
AL
Magro
Angelo J
AJ
Braz
Antonio S K
AS
Fontes
Marcos R M
MR
Giglio
José R
JR
Soares
Andreimar M
AM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017837:Bothrops / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D045726:Metalloproteases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010974:Platelet Aggregation / Q000187:drug effects; D010975:Platelet Aggregation Inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013816:Thermodynamics
D003435:Crotalid Venoms; D018076:DNA, Complementary; D010975:Platelet Aggregation Inhibitors; D045726:Metalloproteases