Molecular characterization and phylogenetic analysis of BjussuMP-I: a RGD-P-III class hemorrhagic metalloprotease from Bothrops jararacussu snake venom.
Snake venom metalloproteases (SVMPs) embody zinc-dependent multidomain enzymes responsible for a relevant pathophysiology in envenomation, including local and systemic hemorrhage. The molecular features responsible for hemorrhagic potency of SVMPs have been associated with their multidomains structures which can target these proteins them to several receptors of different tissues and cellular types. BjussuMP-I, a SVMP isolated from the Bothrops jararacussu venom, has been characterized as a P-III hemorrhagic metalloprotease. The complete cDNA sequence of BjussuMP-I with 1641bp encodes open reading frames of 547 amino acid residues, which conserve the common domains of P-III high molecular weight hemorrhagic metalloproteases: (i) pre-pro-peptide, (ii) metalloprotease, (iii) disintegrin-like and (iv) rich cysteine domain. BjussuMP-I induced lyses in fibrin clots and inhibited collagen- and ADP-induced platelet aggregation. We are reporting, for the first time, the primary structure of an RGD-P-III class snake venom metalloprotease. A phylogenetic analysis of the BjussuMP-I metalloprotease/catalytic domain was performed to get new insights into the molecular evolution of the metalloproteases. A theoretical molecular model of this domain was built through folding recognition (threading) techniques and refined by molecular dynamics simulation. Then, the final BjussuMP-I catalytic domain model was compared to other SVMPs and Reprolysin family proteins in order to identify eventual structural differences, which could help to understand the biochemical activities of these enzymes. The presence of large hydrophobic areas and some conserved surface charge-positive residues were identified as important features of the SVMPs and other metalloproteases.
Mazzi
Maurício V
MV
Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, FCFRP, Universidade de São Paulo, USP, Ribeirão Preto, SP, Brazil.
Magro
Angelo J
AJ
Amui
Saulo F
SF
Oliveira
Clayton Z
CZ
Ticli
Fábio K
FK
Stábeli
Rodrigo G
RG
Fuly
André L
AL
Rosa
José C
JC
Braz
Antônio S K
AS
Fontes
Marcos R M
MR
Sampaio
Suely V
SV
Soares
Andreimar M
AM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017837:Bothrops / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D020134:Catalytic Domain / Q000235:genetics; D003198:Computer Simulation; D003435:Crotalid Venoms / Q000737:chemistry* / Q000145:classification / Q000235:genetics* / Q000633:toxicity; D018076:DNA, Complementary / Q000235:genetics; D005342:Fibrinolysis / Q000187:drug effects; D066298:In Vitro Techniques; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000145:classification / Q000235:genetics* / Q000633:toxicity; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D010974:Platelet Aggregation / Q000187:drug effects; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D055672:Static Electricity; D013816:Thermodynamics
D003435:Crotalid Venoms; D018076:DNA, Complementary; C492638:BjussuMP-I; D008666:Metalloendopeptidases