Molecular and functional characterization of a new non-hemorrhagic metalloprotease from Bothrops jararacussu snake venom with antiplatelet activity

• Published in: Peptides (New York, N.Y. : 1980) Vol. 28; no. 12; pp. 2328 - 2339
• Main Authors: Marcussi, Silvana, Bernardes, Carolina P., Santos-Filho, Norival A., Mazzi, Maurício V., Oliveira, Clayton Z., Izidoro, Luiz Fernando M., Fuly, André L., Magro, Angelo J., Braz, Antonio S.K., Fontes, Marcos R.M., Giglio, José R., Soares, Andreimar M.
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01-12-2007; Elsevier Science
• Subjects: Amino Acid Sequence; Animals; Antiplatelet activity; Base Sequence; Biological activity; Biological and medical sciences; Bothrops - genetics; Bothrops - metabolism; Bothrops jararacussu; cDNA; Cloning, Molecular; Crotalid Venoms - chemistry; Crotalid Venoms - enzymology; Crotalid Venoms - genetics; Crotalid Venoms - pharmacology; DNA, Complementary - genetics; Fibrinogenase; Fundamental and applied biological sciences. Psychology; Humans; In Vitro Techniques; Metalloprotease; Metalloproteases - chemistry; Metalloproteases - genetics; Metalloproteases - isolation & purification; Metalloproteases - metabolism; Models, Molecular; Molecular model; Molecular Sequence Data; Platelet Aggregation - drug effects; Platelet Aggregation Inhibitors - chemistry; Platelet Aggregation Inhibitors - isolation & purification; Protein Conformation; Sequence Homology, Amino Acid; Snake venom; Substrate Specificity; Thermodynamics; Vertebrates: endocrinology; BjussuMP-II; PBS; Bothrops jararacussu; Snake venom; MP; Fibrinogenase; CK; MHD; Metalloprotease; BjussuMP-I; EDTA; Biological activity; Antiplatelet activity; EGTA; CT; NADH; TCA; SVMPs; RCF; RT-PCR; Molecular model; cDNA; TEMED; Metalloprotease,Fibrinogenase,Snake venom,Bothrops jararacussu,Biological activity,Antiplatelet activity,cDNA,Molecular model; Enzyme; Metalloendopeptidases; Hemorrhage; Antiplatelet agent; Ophidia; Characterization; Peptidases; Vertebrata; Venom; Hydrolases; Reptilia
• ISSN: 0196-9781; 1873-5169
• DOI: 10.1016/j.peptides.2007.10.010

BjussuMP-II is an acidic low molecular weight metalloprotease (
Mr
∼
24,000 and pI
∼
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 615
bp 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.