Amino acid sequence and crystal structure of BaP1, a metalloproteinase from Bothrops asper snake venom that exerts multiple tissue-damaging activities

Department of Physics, IBILCE/UNESP, CP 136, Sao José de Rio Preto, CEP 15054-000, Brazil.
Protein Science (Impact Factor: 2.85). 11/2003; 12(10):2273-81. DOI: 10.1110/ps.03102403
Source: PubMed

ABSTRACT BaP1 is a 22.7-kD P-I-type zinc-dependent metalloproteinase isolated from the venom of the snake Bothrops asper, a medically relevant species in Central America. This enzyme exerts multiple tissue-damaging activities, including hemorrhage, myonecrosis, dermonecrosis, blistering, and edema. BaP1 is a single chain of 202 amino acids that shows highest sequence identity with metalloproteinases isolated from the venoms of snakes of the subfamily Crotalinae. It has six Cys residues involved in three disulfide bridges (Cys 117-Cys 197, Cys 159-Cys 181, Cys 157-Cys 164). It has the consensus sequence H(142)E(143)XXH(146)XXGXXH(152), as well as the sequence C(164)I(165)M(166), which characterize the "metzincin" superfamily of metalloproteinases. The active-site cleft separates a major subdomain (residues 1-152), comprising four alpha-helices and a five-stranded beta-sheet, from the minor subdomain, which is formed by a single alpha-helix and several loops. The catalytic zinc ion is coordinated by the N(epsilon 2) nitrogen atoms of His 142, His 146, and His 152, in addition to a solvent water molecule, which in turn is bound to Glu 143. Several conserved residues contribute to the formation of the hydrophobic pocket, and Met 166 serves as a hydrophobic base for the active-site groups. Sequence and structural comparisons of hemorrhagic and nonhemorrhagic P-I metalloproteinases from snake venoms revealed differences in several regions. In particular, the loop comprising residues 153 to 176 has marked structural differences between metalloproteinases with very different hemorrhagic activities. Because this region lies in close proximity to the active-site microenvironment, it may influence the interaction of these enzymes with physiologically relevant substrates in the extracellular matrix.

Download full-text


Available from: José María Gutiérrez, Sep 27, 2015
22 Reads
  • Source
    • "The crystal structures for various SVMPs of the P-I class have been reported and showed remarkable similarity in terms of folding, despite the fact that there are different disulfide bond patterns observed among these SVMPs (Gomis-Ruth et al., 1993; Huang et al., 2002; Watanabe et al., 2003; Zhang et al., 1994). Interestingly, the determination of crystal structures of P-III class SVMPs also showed that the folding of the metalloproteinase domain is similar between these enzymes and those of the P-I class (Igarashi et al., 2007; Muniz et al., 2008; Takeda et al., 2006, 2007). "
    [Show abstract] [Hide abstract]
    ABSTRACT: By catalyzing limited proteolysis or extensive degradation, proteolytic enzymes determine the fate of most proteins in an organism. In the evolutionary process of snake venoms, genes encoding proteinases were tailored to generate potent toxins to target key physiological proteins and thereby play a critical role in prey capture, immobilization and defense against predators. In Bothrops jararaca, metalloproteinases and serine proteinases are among the most abundant toxins both in newborn and adult venoms. In this review, we examine the proteinase-rich venom proteome of B. jararaca and how the proteinases act in a complex and heterogeneous fashion to exert their deleterious local and systemic effects.
    Toxin Reviews 06/2014; 33(4). DOI:10.3109/15569543.2014.922581 · 0.65 Impact Factor
  • Source
    • "leucurolysin-a from B. leucurus (see [32], P84907.2); Bap1 from B. asper (see [33–35], 1ND1_A); BaTX-I from B. atrox (see [36], P0DJE1.1); BnP1 from Bothropoides pauloensis (see [37], P0C6S0.1). "
    [Show abstract] [Hide abstract]
    ABSTRACT: The present study aimed to evaluate the proteolytic and biological activities of a new metalloproteinase from B. moojeni venom. The purification of BmooMP α -II was carried out through two chromatographic steps (ion-exchange and affinity). BmooMP α -II is a monomeric protein with an apparent molecular mass of 22.5 kDa on SDS-PAGE 14% under nonreducing conditions. The N-terminal sequence (FSPRYIELVVVADHGMFTKYKSNLN) revealed homology with other snake venom metalloproteinases, mainly among P-I class. BmooMP α -II cleaves A α -chain of fibrinogen followed by B β -chain, and does not show any effect on the γ -chain. Its optimum temperature and pH for the fibrinogenolytic activity were 30-50°C and pH 8, respectively. The inhibitory effects of EDTA and 1,10-phenantroline on the fibrinogenolytic activity suggest that BmooMP α -II is a metalloproteinase. This proteinase was devoid of haemorrhagic, coagulant, or anticoagulant activities. BmooMP α -II caused morphological alterations in liver, lung, kidney, and muscle of Swiss mice. The enzymatically active protein yet inhibited collagen, ADP, and ristocetin-induced platelet aggregation in a concentration-dependent manner. Our results suggest that BmooMP α -II contributes to the toxic effect of the envenomation and that more investigations to elucidate the mechanisms of inhibition of platelet aggregation may contribute to the studies of snake venom on thrombotic disorders.
    BioMed Research International 06/2014; 2014:352420. DOI:10.1155/2014/352420 · 3.17 Impact Factor
  • Source
    • "Furthermore, by comparing the crystal structure of Bap1 (a P-I SVMP from Bothrops asper) with other hemorrhagic and non-hemorrhagic PI SVMPs [163], the authors of this study showed that an interconnecting loop, formed by amino residues 153–176, show significant structural deviations among SVMPs. This interconnecting loop lies close to the catalytic active-site and may influence the interaction of the SVMPs with extracellular proteins of the basement membrane of capillaries [163]. Another study that employed several bioinformatics and proteomics tools successfully classified up to 19 hemorrhagic and non-hemorrhagic PI metalloproteinases on the basis of their total polar molecular surface area. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Snake venom toxins are responsible for causing severe pathology and toxicity following envenomation including necrosis, apoptosis, neurotoxicity, myotoxicity, cardiotoxicity, profuse hemorrhage, and disruption of blood homeostasis. Clinically, snake venom toxins therefore represent a significant hazard to snakebite victims which underscores the need to produce more efficient anti-venom. Some snake venom toxins, however, have great potential as drugs for treating human diseases. In this review, we discuss the biochemistry, structure/function, and pathology induced by snake venom toxins on human tissue. We provide a broad overview of cobra venom cytotoxins, catalytically active and inactive phospholipase A2s (PLA2s), and Zn2+-dependent metalloproteinases. We also propose biomedical applications whereby snake venom toxins can be employed for treating human diseases. Cobra venom cytotoxins, for example, may be utilized as anti-cancer agents since they are efficient at destroying certain types of cancer cells including leukemia. Additionally, increasing our understanding of the molecular mechanism(s) by which snake venom PLA2s promote hydrolysis of cell membrane phospholipids can give insight into the underlying biomedical implications for treating autoimmune disorders that are caused by dysregulated endogenous PLA2 activity. Lastly, we provide an exhaustive overview of snake venom Zn2+-dependent metalloproteinases and suggest ways by which these enzymes can be engineered for treating deep vein thrombosis and neurodegenerative disorders.
    01/2014; 4(1):1000181. DOI:10.4172/2161-0495.1000181
Show more