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Snake venom thrombin-like enzyme

Roney Vander Dos Santos, Fabian Villalta-Romero, Danijela Stanisic, Luiz Borro, Goran Neshich, Ljubica Tasic
Around 5.5 million people suffer from snakebites per year, with about 400,000 cases with some type of sequelae, such as amputation, and 20,000 to 125,000 cases with the fatal end. Usually, the victim outcome depends on correct, agile and many times in situ intervention based on the proper identification of the snake venom type and its potential effects, among other factors. Therefore, knowledge on the snake venom composition and a research on inhibitors of snake venom target components might ameliorate envenoming dangerous outcome...
January 11, 2018: Toxicon: Official Journal of the International Society on Toxinology
Patrícia Rodrigues Orsi, Fernanda Cruz Landim-Alvarenga, Luis Antônio Justulin, Ramon Kaneno, Marjorie de Assis Golim, Daniela Carvalho Dos Santos, Camila Fernanda Zorzella Creste, Eunice Oba, Leandro Maia, Benedito Barraviera, Rui Seabra Ferreira
BACKGROUND: The injection of mesenchymal stem cells (MSCs) directly into the bone of osteoporotic (OP) patients for rapid recovery has been studied worldwide. Scaffolds associated with MSCs are used to maintain and avoid cell loss after application. A unique heterologous fibrin sealant (HFS) derived from snake venom was evaluated for the cytotoxicity of its main components and as a three-dimensional biological scaffold for MSCs to repair a critical femur defect in osteoporotic rats. METHODS: The cytotoxicity of HFS was assessed using a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) assay and transmission electron microscopy...
September 29, 2017: Stem Cell Research & Therapy
An Li, Chuan Zhang, Jie Wang, Junjie Wang, Hailong Jiang, Jianzhong Li, Xingyuan Ma, Wen Zhang, Yiming Lu
OBJECTIVES: To identify a new member of serine proteases from Deinagkistrodon acutus via phage display technique and appraise its biocatalytic activities. RESULTS: A novel thrombin-like enzyme gene was cloned by screening the phage display library of D. acutus venom gland. The gene has a 783 bp ORF encoding 260 amino acids. A recombinant enzyme expression vector was constructed and the fused protein was expressed in Escherichia coli. The protein was purified showing a single band of approx...
September 21, 2017: Biotechnology Letters
Vance G Nielsen
While snake venom derived enzymes, such as the thrombin-like activity possessing ancrod, have been used to treat thrombotic disease by defibrinogenating patients, the therapeutic potential of fibrinogenolytic snake venom enzymes, such as those derived from Crotalus atrox, have not been fully explored. However, one of the potential risks of administering fibrinogenolytic enzymes to effect defibrinogenation is hemorrhage secondary to hypofibrinogenemia. The present investigation sought to determine if human fibrinogen modified with carbon monoxide (CO) and iron (Fe) could resist degradation by C...
November 2017: Journal of Thrombosis and Thrombolysis
S Rahim
BACKGROUND: Literature supports that very rarely the venom of "Hemotoxic Snake" (Russell's Vipers) affect haemostasis by secreting thrombin-like enzyme which promotes formation of unstable clots. Such clots could lead to fibrin deposition in micro circulation that in turn consume platelets and coagulation factors (consumption coagulopathy). Clinical presentation is mainly spontaneous bleeding and coagulopathy. In this case the significant effects of altered coagulation were observed in retropetitoneum and the patient presented in casuality as acute abdomen...
September 2017: South African Journal of Surgery. Suid-Afrikaanse Tydskrif Vir Chirurgie
Rui Seabra Ferreira, Luciana Curtolo de Barros, Luciana Patrícia Fernandes Abbade, Silvia Regina Catharino Sartori Barraviera, Maria Regina Cavariani Silvares, Leticia Gomes de Pontes, Lucilene Delazari Dos Santos, Benedito Barraviera
Hemostatic and adhesive agents date back to World War II, when homologous fibrin sealant came onto scene. Considering that infectious diseases can be transmitted via human blood, a new heterologous fibrin sealant was standardized in the 1990s. Its components were a serine protease (a thrombin-like enzyme) extracted from the venom of Crotalus durissus terrificus snakes and a fibrinogen-rich cryoprecipitate extracted from the blood of Bubalus bubalis buffaloes. This new bioproduct has been used as a coagulant, sealant, adhesive and recently as a candidate scaffold for mesenchymal stem cells and bone and cartilage repair...
2017: Journal of Venomous Animals and Toxins Including Tropical Diseases
Natalia Perussi Biscola, Luciana Politti Cartarozzi, Suzana Ulian-Benitez, Roberta Barbizan, Mateus Vidigal Castro, Aline Barroso Spejo, Rui Seabra Ferreira, Benedito Barraviera, Alexandre Leite Rodrigues Oliveira
Lesions to the nervous system often produce hemorrhage and tissue loss that are difficult, if not impossible, to repair. Therefore, scar formation, inflammation and cavitation take place, expanding the lesion epicenter. This significantly worsens the patient conditions and impairment, increasing neuronal loss and glial reaction, which in turn further decreases the chances of a positive outcome. The possibility of using hemostatic substances that also function as a scaffold, such as the fibrin sealant, reduces surgical time and improve postoperative recovery...
2017: Journal of Venomous Animals and Toxins Including Tropical Diseases
Ok Hwan Kim, Kil-Sang Cho, Young Seomun, Jong-Tak Kim, Kwang-Hoe Chung
Recombinant batroxobin is a thrombin-like enzyme of Bothrops atrox moojeni venom. To evaluate its toxicological effect, it was highly expressed in Pichia pastorisand successfully purified to homogeneity from culture broth supernatant following Good Manufacturing Practice (GMP). The maximum tolerated dose of the recombinant batroxobin was examined in Sprague-Dawley (SD) rat and Beagle dogs following Good Laboratory Practice (GLP) regulations. The approximate lethal dose of recombinant batroxobin was 10 National Institute of Health (NIH) u/kg in male and female rats...
April 2017: Toxicon: Official Journal of the International Society on Toxinology
Vance G Nielsen, Charles M Bazzell
A complication of defibrinogenation therapy with snake venom enzymes such as ancrod is hypofibrinogenemia associated bleeding secondary to no human-derived inhibitor being available to inactivate or diminish the activity of such enzymes. Of interest, ancrod contains a critical histidine residue without which enzymatic activity is inhibited, and carbon monoxide has been demonstrated to inhibit biomolecular function by interacting with histidine moieties in ion channels. We tested the hypothesis that exposure of three different snake venoms containing serine proteases with thrombin-like activity (which included ancrod) to carbon monoxide derived from carbon monoxide releasing molecule-2 would diminish their effects on plasmatic coagulation as assessed by thrombelastography...
February 2017: Journal of Thrombosis and Thrombolysis
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+)...
November 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
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