{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,11]],"date-time":"2026-10-11T13:23:02Z","timestamp":1791724982897,"version":"4.3.4"},"reference-count":37,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2013,12,4]],"date-time":"2013-12-04T00:00:00Z","timestamp":1386115200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>In this work, we describe the molecular cloning and pharmacological properties of an acidic phospholipase A2 (PLA2) isolated from Bothrops pauloensis snake venom. This enzyme, denominated BpPLA2-TXI, was purified by four chromatographic steps and represents 2.4% of the total snake venom protein content. BpPLA2-TXI is a monomeric protein with a molecular mass of 13.6 kDa, as demonstrated by Matrix-assisted laser desorption\/ionization time-of-flight mass spectrometry (MALDI-TOF) analysis and its theoretical isoelectric point was 4.98. BpPLA2-TXI was catalytically active and showed some pharmacological effects such as inhibition of platelet aggregation induced by collagen or ADP and also induced edema and myotoxicity. BpPLA2-TXI displayed low cytotoxicity on TG-180 (CCRF S 180 II) and Ovarian Carcinoma (OVCAR-3), whereas no cytotoxicity was found in regard to MEF (Mouse Embryonic Fibroblast) and Sarcoma 180 (TIB-66). The N-terminal sequence of forty-eight amino acid residues was determined by Edman degradation. In addition, the complete primary structure of 122 amino acids was deduced by cDNA from the total RNA of the venom gland using specific primers, and it was significantly similar to other acidic D49 PLA2s. The phylogenetic analyses showed that BpPLA2-TXI forms a group with other acidic D49 PLA2s from the gender Bothrops, which are characterized by a catalytic activity associated with anti-platelet effects.<\/jats:p>","DOI":"10.3390\/toxins5122403","type":"journal-article","created":{"date-parts":[[2013,12,4]],"date-time":"2013-12-04T12:02:38Z","timestamp":1386158558000},"page":"2403-2419","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":29,"title":["Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom"],"prefix":"10.3390","volume":"5","author":[{"given":"Francis","family":"Ferreira","sequence":"first","affiliation":[{"name":"Institute of Genetics and Biochemistry, Federal University of Uberl\u00e2ndia, UFU,  38400902 Uberl\u00e2ndia-MG, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M\u00e1rio","family":"Gomes","sequence":"additional","affiliation":[{"name":"Institute of Genetics and Biochemistry, Federal University of Uberl\u00e2ndia, UFU,  38400902 Uberl\u00e2ndia-MG, Brazil"},{"name":"Department of Chemical and Physical, State University of Southwest Bahia (UESB),  45506-210 Jequi\u00e9-BA, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dayane","family":"De Souza","sequence":"additional","affiliation":[{"name":"Institute of Genetics and Biochemistry, Federal University of Uberl\u00e2ndia, UFU,  38400902 Uberl\u00e2ndia-MG, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah","family":"Gimenes","sequence":"additional","affiliation":[{"name":"Institute of Genetics and Biochemistry, Federal University of Uberl\u00e2ndia, UFU,  38400902 Uberl\u00e2ndia-MG, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Let\u00edcia","family":"Castanheira","sequence":"additional","affiliation":[{"name":"Institute of Genetics and Biochemistry, Federal University of Uberl\u00e2ndia, UFU,  38400902 Uberl\u00e2ndia-MG, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M\u00e1rcia","family":"Borges","sequence":"additional","affiliation":[{"name":"Ezequiel Dias Foundation, FUNED, 30510-010 Belo Horizonte-MG, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renata","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"Institute of Genetics and Biochemistry, Federal University of Uberl\u00e2ndia, UFU,  38400902 Uberl\u00e2ndia-MG, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kelly","family":"Yoneyama","sequence":"additional","affiliation":[{"name":"Institute of Genetics and Biochemistry, Federal University of Uberl\u00e2ndia, UFU,  38400902 Uberl\u00e2ndia-MG, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Brandeburgo","sequence":"additional","affiliation":[{"name":"Institute of Genetics and Biochemistry, Federal University of Uberl\u00e2ndia, UFU,  38400902 Uberl\u00e2ndia-MG, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Veridiana","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"Institute of Genetics and Biochemistry, Federal University of Uberl\u00e2ndia, UFU,  38400902 Uberl\u00e2ndia-MG, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2013,12,4]]},"reference":[{"key":"ref_1","first-page":"155","article-title":"Nomes populares e aspectos ecol\u00f3gicos de Bothrops pauloensis (Amaral, 1925) em \u00e1reas antropizadas do Tri\u00e2ngulo e Alto Parana\u00edba, Minas Gerais","volume":"20","author":"Valle","year":"2008","journal-title":"Rev. Bras. Zooci\u00eancias"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2707","DOI":"10.1016\/j.jprot.2012.03.028","article-title":"Combined snake venomics and venom gland transcriptomic analysis of Bothropoides pauloensis","volume":"75","author":"Rodrigues","year":"2012","journal-title":"J. Proteom."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s10557-008-6132-9","article-title":"Phospholipase A2 Biochemistry","volume":"23","author":"Burke","year":"2009","journal-title":"Cardiovasc. Drugs Ther."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/S0090-6980(02)00020-5","article-title":"Phospholipase A2 enzymes","volume":"69","author":"Kudo","year":"2002","journal-title":"Prostag. Other Lipid Mediat."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1246","DOI":"10.1016\/j.bbalip.2006.07.011","article-title":"The phospholipase A2 superfamily and its group numbering system","volume":"1761","author":"Schaloske","year":"2006","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1194\/jlr.R800033-JLR200","article-title":"Phospholipase A2 structure\/function, mechanism, and signaling","volume":"50","author":"Burke","year":"2009","journal-title":"J. Lipids Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1016\/0041-0101(96)00036-0","article-title":"Phospolipase A2\u2014A structural review","volume":"34","author":"Arni","year":"1996","journal-title":"Toxicon"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1219","DOI":"10.1007\/s00204-011-0665-6","article-title":"Molecular characterization of an acidic phospholipase A2 from Bothrops pirajai snake venom: Synthetic-peptide identifies its anti-platelet region","volume":"85","author":"Teixeira","year":"2011","journal-title":"Arch. Toxicol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.jpba.2012.04.008","article-title":"Isolation and expression of a hypotensive and anti-platelet acidic phospholipase A2 from Bothrops moojeni snake venom","volume":"73","author":"Silveira","year":"2013","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1016\/j.toxicon.2004.06.008","article-title":"Bactericidal and neurotoxic activities of two myotoxic phospholipases A2 from Bothrops neuwiedi pauloensis snake venom","volume":"44","author":"Rodrigues","year":"2004","journal-title":"Toxicon"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1006\/abbi.2000.1790","article-title":"Structural and functional characterization of BnSP-7, a Lys49 myotoxic PLA2 homologue from Bothrops neuwiedi pauloensis venom","volume":"37","author":"Soares","year":"2000","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/j.toxicon.2007.03.005","article-title":"Isolation and functional characterization of a new myotoxic acidic phospholipase A2 from Bothrops pauloensis snake venom","volume":"50","author":"Rodrigues","year":"2007","journal-title":"Toxicon"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.toxicon.2010.02.031","article-title":"Isolation and functional characterization of a new acidic PLA2 Ba SpII RP4 of the Bothrops alternatus snake venom from Argentina","volume":"56","author":"Denegri","year":"2010","journal-title":"Toxicon"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"908","DOI":"10.1016\/j.toxicon.2008.08.020","article-title":"A new acidic myotoxic, anti-platelet and prostaglandin I2 inductor phospholipase A2 isolated from Bothrops moojeni snake venom","volume":"52","author":"Silveira","year":"2008","journal-title":"Toxicon"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1016\/j.bbrc.2003.10.047","article-title":"Crystal structures of BnSP-7 and BnSP-6, two Lys49-phospholipases A2: Quaternary structure and inhibition mechanism insights","volume":"311","author":"Magro","year":"2003","journal-title":"Biochem. Biophys. Res. Comun."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1","DOI":"10.2174\/1385272043369610","article-title":"Phospholipase A2 myotoxins from Bothrops snake venoms: Structure-function relationship","volume":"8","author":"Soares","year":"2004","journal-title":"Curr. Org. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1583","DOI":"10.1016\/j.bbapap.2009.07.005","article-title":"Crystal structure of a phospholipase A2 homologue complexed with p-bromophenacyl bromide reveals important structural changes associated with the inhibition of myotoxic activity","volume":"1794","author":"Fernandes","year":"2009","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_18","first-page":"1","article-title":"The growing phospholipase A2 superfamily of signal transduction enzymes","volume":"22","author":"Dennis","year":"1997","journal-title":"TiBS"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1016\/j.biochi.2009.12.006","article-title":"Isolation of an acidic phospholipase A2 from the venom of the snake Bothrops asper of Costa Rica: Biochemical and toxicological characterization","volume":"92","author":"Angulo","year":"2010","journal-title":"Biochimie"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1016\/S0006-2952(02)01210-8","article-title":"Structural and functional characterization of an acidic platelet aggregation inhibitor and hypotensive phospholipase A2 from Bothrops jararacussu snake venom","volume":"64","author":"Soares","year":"2002","journal-title":"Biochem. Pharmacol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1470","DOI":"10.1016\/S1357-2725(03)00129-8","article-title":"Miotoxicity induced by an acidic Asp-49 phospholipase A2 isolated from Lachesis muta (Bushmaster) snake venom. Comparison with lysophosphatidylcholine","volume":"35","author":"Fuly","year":"2003","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"915","DOI":"10.1016\/j.toxicon.2003.11.005","article-title":"Skeletal muscle degeneration induced by venom phospholipases A2: Insights into the mechanisms of local and systemic myotoxicity","volume":"42","author":"Ownby","year":"2003","journal-title":"Toxicon"},{"key":"ref_23","first-page":"371","article-title":"Bothrops moojeni myotoxin-II, a Lys-49-phospholipase A2 homologue: An example of function versatility of snake venom proteins","volume":"142","author":"Amui","year":"2006","journal-title":"Compar. Biochem. Physiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1645","DOI":"10.1016\/j.peptides.2008.05.021","article-title":"Myotoxic phospholipase A2 isolated from Bothrops brazili snake venom and synthetic peptides derived from their C-terminal region: Cytotoxic effect on microorganism and tumor cells","volume":"29","author":"Costa","year":"2008","journal-title":"Peptides"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1016\/j.bcp.2006.04.032","article-title":"BE-I-PLA2, a novel acidic phospholipase A2 from Bothrops erythromelas venom: Isolation, cloning and characterization as potent anti-platelet and inductor of prostaglandin I2 release by endothelial cells","volume":"72","author":"Modesto","year":"2006","journal-title":"Biochem. Pharmacol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"5605","DOI":"10.1073\/pnas.92.12.5605","article-title":"Accelerated evolution in the protein-coding regions is universal in crotalinae snake venom gland phospholipase A2 isozyme genes","volume":"92","author":"Nakashima","year":"1995","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1042\/BJ20040125","article-title":"Molecular evolution and structure-activity relationships of crotoxin-B like asparagin 6 phospholipase A2 in pit viper venom","volume":"381","author":"Chen","year":"2004","journal-title":"Biochem. J."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"672","DOI":"10.1016\/j.toxicon.2006.06.021","article-title":"Discovery of novel (Arg49) phospholipase A2 isoenzymes from ProtoBothrops elegans venom and regional evolution of Crotalinae snake venom phospholipase A2 isoenzymes in the southwestern islands of Japan and Taiwan","volume":"48","author":"Chijiwa","year":"2006","journal-title":"Toxicon"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1007\/s00239-008-9067-7","article-title":"Rapid evolution by positive selection and gene gain and loss: PLA2 venom genes in closely related Sistrurus rattlesnakes with divergent diets","volume":"66","author":"Gibbs","year":"2008","journal-title":"J. Mol. Evol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1016\/S0003-9861(02)00747-6","article-title":"Geographic variations, cloning and functional analyses of the venom acidic phospholipases A2 of Crotalus viridis viridis","volume":"411","author":"Tsai","year":"2003","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.cbpb.2011.10.002","article-title":"Biochemical and enzymatic characterization of BpMP-I, a fibrinogenolytic metalloproteinase isolated from Bothropoides. pauloensis snake venom","volume":"161","author":"Naves","year":"2012","journal-title":"Comp. Biochem. Physiol."},{"key":"ref_32","unstructured":"Ferreira, F.B. (2011). Unpublished work."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1038\/227680a0","article-title":"Cleavage of structural proteins during the assembly of the head of bacteriophage T4","volume":"227","author":"Laemmli","year":"1970","journal-title":"Nature"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/0005-2744(68)90248-9","article-title":"Purification and properties of phospholipase A2 from porcine pancreas","volume":"159","author":"Postema","year":"1968","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1016\/0041-0101(88)90010-4","article-title":"Alternative in vitro method for testing the potency of the polyvalent antivenom produced in Costa Rica","volume":"26","author":"Gutierrez","year":"1988","journal-title":"Toxicon"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1016\/0041-0101(92)90534-C","article-title":"Gene expression in Echis carinatus (carpet viper) venom glands following milking","volume":"30","author":"Paine","year":"1992","journal-title":"Toxicon"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1093\/sysbio\/sys029","article-title":"MrBayers 3.2: Efficient Bayesian phylogenetic inference and model choice across a large model space","volume":"3","author":"Ronquist","year":"2012","journal-title":"Syst. Biol."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/5\/12\/2403\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:51:08Z","timestamp":1760219468000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/5\/12\/2403"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,12,4]]},"references-count":37,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2013,12]]}},"alternative-id":["toxins5122403"],"URL":"https:\/\/doi.org\/10.3390\/toxins5122403","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,12,4]]}}}