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Snake venom thrombin-like enzyme

Huiqiong Xie, Miao Huang, Qiping Hu, Kejian Sun, Huayu Wu, Wei Shu, Xiaolong Li, Ling Fang
In previous work, a snake venom arginine esterase (SVAE), agkihpin from the venom of Gloydius halys Pallas, was isolated and its biochemical data including Mr, PI, amino acid components and sugar content was collected. Here, the agkihpin was cloned and further characterized and we found that agkihpin could promote ADP-induced platelets aggregation, hydrolyze fibrin, cleave Aα and Bβ chains of fibrinogen and reduce the thrombosis induced by thrombin. Moreover, agkihpin hydrolyzed TAME with optimum temperatures at 30 °C-45 °C, and the hydrolysis was inhibited by EDTA, PMSF, DTT and promoted by Ca(2+), Fe(3+), Mg(2+), Zn(2+)...
September 22, 2016: Toxicon: Official Journal of the International Society on Toxinology
Thais F Isabel, Guilherme Nunes Moreira Costa, Isabela B Pacheco, Luana G Barbosa, Célio D Santos-Junior, Fernando P P Fonseca, Johara Boldrini França, Flávio Henrique-Silva, Kelly A G Yoneyama, Renata S Rodrigues, Veridiana de Melo Rodrigues
Snake venom serine proteases (SVSPs) are enzymes capable of interfering at several points of hemostasis. Some serine proteases present thrombin-like activity, which makes them targets for the development of therapeutics agents in the treatment of many hemostatic disorders. In this study, a recombinant thrombin-like serine protease, denominated rBpSP-II, was obtained from cDNA of the Bothrops pauloensis venom gland and was characterized enzymatically and biochemically. The enzyme rBpSP-II showed clotting activity on bovine plasma and proteolytic activity on fibrinogen, cleaving exclusively the Aα chain...
June 1, 2016: Toxicon: Official Journal of the International Society on Toxinology
Vance G Nielsen
Fibrinogen depletion via catalysis by snake venom enzymes as a therapeutic strategy to prevent or treat thrombotic disorders was utilized for over four decades, with ancrod being the quintessential agent. However, ancrod eventually was found to not be of clinical utility in large scale stroke trial, resulting in the eventual discontinuation of the administration of the drug for any indication. It was hypothesized that ancrod, possessing thrombin-like activity, may have unappreciated robust coagulation kinetics...
August 2016: Journal of Thrombosis and Thrombolysis
Kayena D Zaqueo, Anderson M Kayano, Thaisa F S Domingos, Laura A Moura, André L Fuly, Saulo L da Silva, Gerardo Acosta, Eliandre Oliveira, Fernando Albericio, Fernando B Zanchi, Juliana P Zuliani, Leonardo A Calderon, Rodrigo G Stábeli, Andreimar M Soares
Snake venom toxins are related not only in detention, death and the promotion of initial digestion of prey but also due to their different biochemical, structural and pharmacological effects they can result in new drugs. Among these toxins snake venom serine proteases (SVSPs) should be highlighted because they are responsible for inducing changes in physiological functions such as blood coagulation, fibrinolysis, and platelet aggregation. This article presents the first serine protease (SP) isolated from Bothrops brazili: BbrzSP-32...
May 2016: Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology
Marcelo Larami Santoro, Thaís do Carmo, Bruna Heloísa Lopes Cunha, André Fonseca Alves, André Zelanis, Solange Maria de Toledo Serrano, Kathleen Fernandes Grego, Savio Stefanini Sant'Anna, Katia Cristina Barbaro, Wilson Fernandes
Lance-headed snakes are found in Central and South America, and they account for most snakebites in Brazil. The phylogeny of South American pitvipers has been reviewed, and the presence of natural and non-natural hybrids between different species of Bothrops snakes demonstrates that reproductive isolation of several species is still incomplete. The present study aimed to analyze the biological features, particularly the thrombin-like activity, of venoms from hybrids born in captivity, from the mating of a female Bothrops erythromelas and a male Bothrops neuwiedi, two species whose venoms are known to display ontogenetic variation...
2015: PloS One
Débora da-Silva-Freitas, Johara Boldrini-França, Eliane Candiani Arantes
PEGylation is considered a successful technique to enhance the therapeutic and biotechnological potentials of peptides, proteins, toxins and drugs. The conjugation of polyethylene glycol (PEG) increases the size and molecular weight of conjugated molecule and improves its pharmacokinetics and pharmacodinamics by increasing water solubility, protecting from enzymatic degradation, reducing renal clearance and limiting immunogenic and antigenic reactions. These features are very useful for therapeutic proteins, since PEGylated proteins exhibit high stability and very low immunogenicity, ensuring a sustained clinical response with minimal dose and less frequent administration...
2015: Protein and Peptide Letters
Maitreyee Sharma, Diganta Das, Janaki Krishnamoorthy Iyer, R Manjunatha Kini, Robin Doley
Composition of Indian Russell's viper (Daboia russelii russelii) venom, a medically important snake and member of "Big Four" snakes of India was done by gel filtration chromatography followed by tandem mass spectrometry. The MS/MS analyses of tryptic digested gel filtration peaks divulged the presence of 63 different proteins belonging to 12 families. Phospholipase A2 (PLA2), serine proteases, metalloproteases, cysteine-rich secretory proteins, l-amino acid oxidase, C-type lectin-like proteins, kunitz-type serine protease inhibitor, disintegrin, nucleotidase, phosphodiesterase, vascular endothelial growth factor and vascular nerve growth factor families were identified...
December 1, 2015: Toxicon: Official Journal of the International Society on Toxinology
Luana V Senise, Karine M Yamashita, Marcelo L Santoro
To attain fully functional biological activity, vitamin-K dependent coagulation factors (VKDCF) are γ-carboxylated prior to secretion from liver. Warfarin impairs the γ-carboxylation, and consequently their physiological function. Bothrops jararaca snake venom (BjV) contains several activators of blood coagulation, especially procoagulant enzymes (prothrombin and factor X activators) and thrombin-like enzymes. In order to clarify the relative contribution of prothrombin and factor X activators to the hemostatic disturbances occurring during experimental B...
November 2015: Experimental Biology and Medicine
Kadi-Saci Amel, Laraba-Djebari Fatima
Snake venom serine proteinases (SVSPs) affect various physiological functions including blood coagulation, fibrinolysis, and platelet aggregation. Coagulant serine proteinase (VLCII) was purified from Vipera lebetina venom using three chromatographic steps: gel filtration on SephadexG-75, DEAE-Sephadex A-50, and reversed-phase high-performance liquid chromatography (RP-HPLC) on C8 column. VLCII appeared homogenous (60 kDa) when tested on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). VLCII as a thrombin-like enzyme was able to hydrolyze Nα-CBZ L-arginine-p-nitroanilide hydrochloride and could be a serine protease because it is inhibited by phenylmethylsulfonyl fluoride...
August 2015: Journal of Biochemical and Molecular Toxicology
Paweena Pradniwat, Ponlapat Rojnuckarin
Pit viper venoms can decrease fibrinogen levels in snakebite patients. Studies have shown that the hypofibrinogenemia is a consequence of snake venom thrombin-like enzymes (TLEs), the serine proteases that have the potential to be both diagnostic and therapeutic agents. Exosites of thrombin are the molecular regions that determine the substrate specificities, but its presence and significance in TLEs are unclear. Therefore, the putative exosites of recombinant TLEs derived from Green pit viper (Trimeresurus albolabris), GPV-TL1 and GPV-TL2, were mutated in a Pichia pastoris system...
June 15, 2015: Toxicon: Official Journal of the International Society on Toxinology
Edson T Yamashiro, Ana K Oliveira, Eduardo S Kitano, Milene C Menezes, Inácio L Junqueira-de-Azevedo, Adriana F Paes Leme, Solange M T Serrano
Snake venoms contain serine proteinases that are functionally similar to thrombin and specifically cleave fibrinogen to convert it into fibrin or activate platelets to aggregation. PA-BJ is a serine proteinase from Bothrops jararaca venom that promotes platelet aggregation and this effect is mediated by the G-coupled protein receptors PAR1 and PAR4. In this study we describe an improved procedure to obtain PA-BJ from B. jararaca venom that uses less chromatographic steps, and, interestingly, results in the isolation of eight proteoforms showing slightly different pIs and molecular masses due to variations in their glycosylation levels...
December 2014: Biochimica et Biophysica Acta
Juliana Félix-Silva, Thiago Souza, Yamara A S Menezes, Bárbara Cabral, Rafael B G Câmara, Arnóbio A Silva-Junior, Hugo A O Rocha, Ivanise M M Rebecchi, Silvana M Zucolotto, Matheus F Fernandes-Pedrosa
Snakebites are a serious public health problem due their high morbi-mortality. The main available specific treatment is the antivenom serum therapy, which has some disadvantages, such as poor neutralization of local effects, risk of immunological reactions, high cost and difficult access in some regions. In this context, the search for alternative therapies is relevant. Therefore, the aim of this study was to evaluate the antiophidic properties of Jatropha gossypiifolia, a medicinal plant used in folk medicine to treat snakebites...
2014: PloS One
Ashis K Mukherjee
Proteases from Russell's viper venom (RVV) induce a variety of toxic effects in victim. Therefore, four new RVV protease isoenzymes of molecular mass 32901.044 Da, 333631.179 Da, 333571.472 Da, and 34594.776 Da, were characterized in this study. The first 10 N-terminal residues of these serine protease isoenzymes showed significant sequence homology with N-terminal sequences of snake venom thrombin-like and factor V-activating serine proteases, which was reconfirmed by peptide mass fingerprinting analysis. These proteases were found to be different from previously reported factor V activators isolated from snake venoms...
2014: PloS One
Xiang-Jun Zhou, Jie Zhang, Ci Zhang, Chang-Geng Xu
OBJECTIVE: The aim of this study was to determine the feasibility of using snake venom thrombin-like enzyme (SVTLE) and/or ethylenediaminetetraacetic acid (EDTA) to dissolve calcium oxalate stones in vitro. METHODS: Seven calcium oxalate stones were incubated with various chemolytic agents [EDTA, Tris-HCl/EDTA (TE) buffer or SVTLE diluted in TE buffer]. The pH, calcium concentration, stone weight and stone surface integrity were recorded, as well as related pathological changes to bladder mucosae...
2014: Urologia Internationalis
Frank Denis Torres-Huaco, Cláudio C Werneck, Cristina Pontes Vicente, Talita Vassequi-Silva, Ana Cláudia Coelho Nery-Diez, Camila B Mendes, Edson Antunes, Sérgio Marangoni, Daniela C S Damico
We report a rapid purification method using one-step chromatography of SVSP Rhombeobin (LMR-47) from Lachesis muta rhombeata venom and its procoagulant activities and effects on platelet aggregation. The venom was fractionated by a single chromatographic step in RP-HPLC on a C8 Discovery BIO Wide Pore, showing high degree of molecular homogeneity with molecular mass of 47035.49 Da. Rhombeobin showed amidolytic activity upon BA ρ NA, with a broad optimum pH (7-10) and was stable in solution up to 60°C. The amidolytic activity was inhibited by serine proteinase inhibitors and reducing agents, but not chelating agents...
2013: BioMed Research International
K Maduwage, F E Scorgie, A Silva, S Shahmy, F Mohamed, C Abeysinghe, H Karunathilake, L F Lincz, C A Gnanathasan, G K Isbister
CONTEXT: Limited information exists on the coagulopathy caused by hump-nosed pit viper (Hypnale hypnale) envenoming. OBJECTIVES: This study aimed to characterise the coagulopathy in hump-nosed pit viper bites by measuring laboratory clotting times and factor studies. MATERIALS AND METHODS: Cases of hump-nosed pit viper envenoming were included from a prospective cohort study of Sri Lankan snake-bite patients. Patient age, sex, snake identification, time of bite and clinical effects were recorded...
August 2013: Clinical Toxicology
Dan E Vivas-Ruiz, Gustavo A Sandoval, Julio Mendoza, Rosalina R Inga, Silea Gontijo, Michael Richardson, Johannes A Eble, Armando Yarleque, Eladio F Sanchez
The thrombin-like enzyme from Bothrops barnetti named barnettobin was purified. We report some biochemical features of barnettobin including the complete amino acid sequence that was deduced from the cDNA. Snake venom serine proteases affect several steps of human hemostasis ranging from the blood coagulation cascade to platelet function. Barnettobin is a monomeric glycoprotein of 52 kDa as shown by reducing SDS-PAGE, and contains approx. 52% carbohydrate by mass which could be removed by N-glycosidase. The complete amino acid sequence was deduced from the cDNA sequence...
July 2013: Biochimie
Ying Zheng, Feng-Ping Ye, Jie Wang, Guo-Yang Liao, Yun Zhang, Quan-Shui Fan, Wen-Hui Lee
A serine protease termed Da-36 was isolated from crude venom of Deinagkistrodon acutus. The enzyme was a single chain protein with an apparent molecular weight of 36,000 on SDS-PAGE with an isoelectric point of 6.59. Da-36 could clot human plasma by cleaving the Aα, Bβ and γ chains of fibrinogen and also exhibited arginine esterase activity. The proteolytic activity of Da-36 toward TAME was strongly inhibited by PMSF and moderately affected by benzamidine and aprotinin, indicating that it was a serine protease...
June 1, 2013: Toxicon: Official Journal of the International Society on Toxinology
Anthony J Saviola, David Chiszar, Chardelle Busch, Stephen P Mackessy
BACKGROUND: Vertebrate predators use a broad arsenal of behaviors and weaponry for overcoming fractious and potentially dangerous prey. A unique array of predatory strategies occur among snakes, ranging from mechanical modes of constriction and jaw-holding in non-venomous snakes, to a chemical means, venom, for quickly dispatching prey. However, even among venomous snakes, different prey handling strategies are utilized, varying from the strike-and-hold behaviors exhibited by highly toxic elapid snakes to the rapid strike-and-release envenomation seen in viperid snakes...
2013: BMC Biology
Song-Shan Tang, Xiao-Hua Wang, Juan-Hui Zhang, Bo-Shan Tang, Li Qian, Pei-Ying Li, Lie-Wei Luo
A number of snake venom thrombin-like enzymes (TLEs) have already been characterized. Some TLEs play significant roles in vessel injury hemostasis. A novel TLE (Agacutase) was purified from Deinagkistrodon acutus snake venom by the means of Sephadex G-75, DEAE-Sepharose FF, and Sephadex G-25 column chromatography. Structural analysis indicated that Agacutase is a single-chain glycoprotein with a molecular mass of 31,084 Da, isoelectric point of 4.38, optimal activity at 37 °C and pH 6.6, sugar content of 7...
April 11, 2013: European Journal of Pharmaceutical Sciences
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