{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T06:12:59Z","timestamp":1772777579867,"version":"3.50.1"},"reference-count":54,"publisher":"Portland Press Ltd.","issue":"1","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2004,7,1]]},"abstract":"<jats:p>Some myotoxic or neurotoxic PLA2s (phospholipases A2) from pit viper venoms contain characteristic N6 substitutions. Our survey of the venoms of more than ten pit viper genera revealed that N6-PLA2s exist only in limited Asian pit vipers of two genera, Protobothrops and Gloydius, and exist as either monomers or the basic subunits of heterodimers in some New World pit vipers. For the newly identified N6-PLA2s, the neuromuscular blocking activities were assayed with the chick biventer cervicis neuromuscular tissue, whereas the increased serum creatine kinase level assessed their myotoxicities. The purified N6-PLA2s from Protobothrops mangshanensis and Gloydius intermedius saxatilis were found to be presynaptic neurotoxins. In contrast, all N6-PLA2s from the venoms of Sistrurus miliarius strackeri, S. m. barbouri, Crotalus viridis viridis, C. lepidus lepidus, Cerrophidion godmani and Bothreichis schlegelii were myotoxins without neurotoxicity even in the presence of crotoxin A. Crotoxin-like complexes were for the first time purified from the venoms of Sitrurus catenatus tergeminus, C. mitchelli mitchelli, C. horridus atricaudatus, C. basiliscus and C. durissus cumanensis. The cDNAs encoding six novel N6-PLA2s and subunits of the crotoxin-like complex from S. c. tergeminus were cloned and fully sequenced. Phylogeny analysis showed that two structural subtypes of N6-PLA2s with either F24 or S24 substitution have been evolved in parallel, possibly descended respectively from species related to present-day Protobothrops and Gloydius. Calmodulin binds all the N6-PLA2s but crotoxin A may inhibit its binding to crotoxin B and to other neurotoxic N6-PLA2s. Structure\u2013activity relationships at various regions of the PLA2 molecules were extensively discussed.<\/jats:p>","DOI":"10.1042\/bj20040125","type":"journal-article","created":{"date-parts":[[2004,6,22]],"date-time":"2004-06-22T13:45:11Z","timestamp":1087911911000},"page":"25-34","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":53,"title":["Molecular evolution and structure\u2013function relationships of crotoxin-like and asparagine-6-containing phospholipases A2 in pit viper venoms"],"prefix":"10.1042","volume":"381","author":[{"given":"Yi-Hsuan","family":"CHEN","sequence":"first","affiliation":[{"name":"Institute of Biological Chemistry, Academia Sinica, POB 23-106, Taipei, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ying-Ming","family":"WANG","sequence":"additional","affiliation":[{"name":"Institute of Biological Chemistry, Academia Sinica, POB 23-106, Taipei, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ming-Jhy","family":"HSEU","sequence":"additional","affiliation":[{"name":"Institute of Biological Chemistry, Academia Sinica, POB 23-106, Taipei, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Inn-Ho","family":"TSAI","sequence":"additional","affiliation":[{"name":"Institute of Biological Chemistry, Academia Sinica, POB 23-106, Taipei, Taiwan"},{"name":"Institute of Biochemical Sciences, National Taiwan University, Taipei, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"288","published-online":{"date-parts":[[2004,6,22]]},"reference":[{"key":"2021112212573632100_B1","first-page":"3","article-title":"The rattlesnake and their venom yield and lethal activity","volume-title":"Rattlesnake Venoms: their Actions and Treatments","author":"Glenn","year":"1982"},{"key":"2021112212573632100_B2","doi-asserted-by":"crossref","first-page":"33","DOI":"10.3109\/15569549709064092","article-title":"Rattlesnake neurotoxins: biochemical and biological aspects","volume":"16","author":"Bieber","year":"1990","journal-title":"J. Toxicol. Toxin Rev."},{"key":"2021112212573632100_B3","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1111\/j.1432-1033.1993.tb17946.x","article-title":"Comparison of crotoxin isoforms reveals that stability of the complex plays a major role in its pharmacological action","volume":"214","author":"Faure","year":"1993","journal-title":"Eur. J. Biochem."},{"key":"2021112212573632100_B4","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1016\/0041-0101(85)90014-5","article-title":"The distribution among ophidian venoms of a toxin isolated from the venom of the Mojave rattlesnake (Crotalus scutulatus scutulatus)","volume":"23","author":"Weinstein","year":"1985","journal-title":"Toxicon"},{"key":"2021112212573632100_B5","first-page":"595","article-title":"Hemorrhagic and mojave toxins in the venoms of the offspring of two Mojave rattlesnakes (Crotalus scutulatus scutulatus)","volume":"106B","author":"Rael","year":"1993","journal-title":"Comp. Biochem. Physiol."},{"key":"2021112212573632100_B6","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/S1096-4959(01)00422-5","article-title":"Mojave rattlesnakes (Crotalus scutulatus scutulatus) lacking the acidic subunit DNA sequence lack Mojave toxin in their venom","volume":"130B","author":"Wooldridge","year":"2001","journal-title":"Comp. Biochem. Physiol."},{"key":"2021112212573632100_B7","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":"Guti\u00e9rrez","year":"2003","journal-title":"Toxicon"},{"key":"2021112212573632100_B8","doi-asserted-by":"crossref","first-page":"576","DOI":"10.2307\/1447591","article-title":"Molecular systematics and biogeographical history of pit vipers as determined by mitochondria ribosomal DNA sequences","volume":"1999","author":"Parkinson","year":"1999","journal-title":"Copeia"},{"key":"2021112212573632100_B9","first-page":"29","article-title":"Molecular biology of snake venom phospholipase A2","volume-title":"Venom Phospholipase A2 Enzyme: Structure, Function and Mechanism","author":"Danse","year":"1997"},{"key":"2021112212573632100_B10","doi-asserted-by":"crossref","first-page":"79","DOI":"10.3109\/15569549709016450","article-title":"Phospholipase A2 of Asian snake venoms","volume":"16","author":"Tsai","year":"1997","journal-title":"J. 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J."},{"key":"2021112212573632100_B13","doi-asserted-by":"crossref","first-page":"546","DOI":"10.1007\/s00239-003-2508-4","article-title":"Interisland mutation of a novel phospholipase A2 from Trimeresurus flavoviridis venom and evolution of Crotalinae group II phospholipases A2","volume":"57","author":"Chijiwa","year":"2003","journal-title":"J. Mol. 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Biochem."},{"key":"2021112212573632100_B18","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1016\/0076-6879(87)55023-6","article-title":"Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction","volume":"155","author":"Mullis","year":"1987","journal-title":"Methods Enzymol."},{"key":"2021112212573632100_B19","article-title":"Molecular Cloning: A Laboratory Manual, 2nd edn","author":"Maniatis","year":"1989"},{"key":"2021112212573632100_B20","first-page":"410","article-title":"The isolated chick biventer cervicis nerve-muscle preparation","volume":"15","author":"Ginsborg","year":"1960","journal-title":"Br. J. 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J. Biochem."},{"key":"2021112212573632100_B25","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1111\/j.1476-5381.1981.tb10448.x","article-title":"A study on the interaction of crotapotin with crotoxin phospholipase A2, notexin and other presynaptic neurotoxins","volume":"73","author":"Chang","year":"1981","journal-title":"Br. J. 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Protein Chem."},{"key":"2021112212573632100_B33","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1006\/abbi.2000.2244","article-title":"Dissociation of enzymatic and pharmacological properties of piratoxins-I and -III, two myotoxic phospholipases A2 from Bothrops pirajai snake venom","volume":"387","author":"Soares","year":"2001","journal-title":"Arch. Biochem. Biophys."},{"key":"2021112212573632100_B34","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/S1357-2725(00)00073-X","article-title":"Cloning and cDNA sequence analysis of Lys49 and Asp49 basic phospholipase A2 myotoxin isoforms from Bothrops asper","volume":"33","author":"Lizano","year":"2001","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"2021112212573632100_B35","first-page":"61","article-title":"Comparison of the biological activities in venoms from three subspecies of the South American rattlesnake (Crotalus durissus terrificus, C. durissus cascavella and C. durissus collilineatus)","volume":"122C","author":"Santoro","year":"1999","journal-title":"Comp. Biochem. Physiol."},{"key":"2021112212573632100_B36","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.1016\/0041-0101(87)90268-6","article-title":"A crotoxin homolog from the venom of the uracoan rattlesnake (Crotalus vegrandis)","volume":"25","author":"Kaiser","year":"1987","journal-title":"Toxicon"},{"key":"2021112212573632100_B37","first-page":"337","article-title":"Regional variation in the presence of canebrake toxin in Crotalus horridus venom","volume":"107C","author":"Glenn","year":"1994","journal-title":"Comp. Biochem. Physiol."},{"key":"2021112212573632100_B38","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1111\/j.1432-1033.1994.tb18978.x","article-title":"The origin of the diversity of crotoxin isoforms in the venom of Crotalus durissus terrificus","volume":"223","author":"Faure","year":"1994","journal-title":"Eur. J. Biochem."},{"key":"2021112212573632100_B39","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/S0041-0101(03)00100-4","article-title":"Structural and biological characterization of a crotapotin isoform isolated from Crotalus durissus cascavella venom","volume":"42","author":"de Oliveira","year":"2003","journal-title":"Toxicon"},{"key":"2021112212573632100_B40","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1023\/A:1015332314389","article-title":"Isolation and preliminary enzymatic characterization of a novel PLA2 from Crotalus durissus collilineatus venom","volume":"21","author":"Ponce-Soto","year":"2002","journal-title":"J. Protein Chem."},{"key":"2021112212573632100_B41","doi-asserted-by":"crossref","first-page":"1252","DOI":"10.1111\/j.1471-4159.1989.tb07422.x","article-title":"Binding of crotoxin, a presynaptic phospholipase A2 neurotoxin, to negatively charged phospholipid vesicles","volume":"53","author":"Radvanyi","year":"1989","journal-title":"J. Neurochem."},{"key":"2021112212573632100_B42","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1111\/j.1470-8744.1994.tb00287.x","article-title":"Isolation and characterization of a toxic phospholipase A2 from the venom of the Taiwan habu (Trimeresurus mucrosquamatus)","volume":"19","author":"Chen","year":"1994","journal-title":"Biotechnol. Appl. 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J."},{"key":"2021112212573632100_B45","doi-asserted-by":"crossref","first-page":"2120","DOI":"10.1074\/jbc.270.5.2120","article-title":"Conversion of bovine pancreatic phospholipase A2 at a single site into a competitor of neurotoxic phospholipase A2 by site-directed mutagenesis","volume":"270","author":"Tzeng","year":"1995","journal-title":"J. Biol. Chem."},{"key":"2021112212573632100_B46","doi-asserted-by":"crossref","first-page":"1591","DOI":"10.1016\/S0041-0101(98)00151-2","article-title":"Effect of site directed mutagenesis on the activity of recombinant trimucrotoxin, a neurotoxic phospholipase from Trimeresurus mucrosquamatus venom","volume":"36","author":"Tsai","year":"1998","journal-title":"Toxicon"},{"key":"2021112212573632100_B47","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1006\/jmbi.1998.1987","article-title":"Crystal structure of agkistrodotoxin, a phospholipase A2-type presynaptic neurotoxin from Agkistrodon halys Pallas","volume":"282","author":"Tang","year":"1998","journal-title":"J. Mol. Biol."},{"key":"2021112212573632100_B48","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1515\/BC.1999.071","article-title":"Molecular mimicry between a monoclonal antibody and one subunit of crotoxin, a heterodimeric phospholipase A2 neurotoxin","volume":"380","author":"Choumet","year":"1999","journal-title":"Biol. Chem."},{"key":"2021112212573632100_B49","first-page":"383","article-title":"The hemolytic site of the basic phospholipase A2 from Agkistrodon halys pallas","volume":"34","author":"Jiao","year":"2002","journal-title":"Acta Biochim. Biophys. 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