title: Novel Catalytically-Inactive PII Metalloproteinases from a Viperid Snake Venom with Substitutions in the Canonical Zinc-Binding Motif.
abstract: Snake venom metalloproteinases (SVMPs) play key biological roles in prey immobilization and digestion. The majority of these activities depend on the hydrolysis of relevant protein substrates in the tissues. Hereby, we describe several isoforms and a cDNA clone sequence, corresponding to PII SVMP homologues from the venom of the Central American pit viperBothriechis lateralis, which have modifications in the residues of the canonical sequence of the zinc-binding motif HEXXHXXGXXH. As a consequence, the proteolytic activity of the isolated proteins was undetectable when tested on azocasein and gelatin. These PII isoforms comprise metalloproteinase and disintegrin domains in the mature protein, thus belonging to the subclass PIIb of SVMPs. PII SVMP homologues were devoid of hemorrhagic and in vitro coagulant activities, effects attributed to the enzymatic activity of SVMPs, but induced a mild edema. One of the isoforms presents the characteristic RGD sequence in the disintegrin domain and inhibits ADP- and collagen-induced platelet aggregation. Catalytically-inactive SVMP homologues may have been hitherto missed in the characterization of snake venoms. The presence of such enzymatically-inactive homologues in snake venoms and their possible toxic and adaptive roles deserve further investigation.
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José 11501, Costa Rica. ecamachoum@gmail.com.
authors.forename: Erika
authors.initials: E
authors.lastname: Camacho
authors.affiliation: Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Científicas, Valencia 46010, Spain. libia.sanz@ibv.csic.es.
authors.forename: Libia
authors.initials: L
authors.lastname: Sanz
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José 11501, Costa Rica. teresa.escalante@ucr.ac.cr.
authors.forename: Teresa
authors.initials: T
authors.lastname: Escalante
authors.affiliation: Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Científicas, Valencia 46010, Spain. aperez@ibv.csic.es.
authors.forename: Alicia
authors.initials: A
authors.lastname: Pérez
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José 11501, Costa Rica. fvillalta85@gmail.com.
authors.forename: Fabián
authors.initials: F
authors.lastname: Villalta
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José 11501, Costa Rica. bruno.lomonte@ucr.ac.cr.
authors.forename: Bruno
authors.initials: B
authors.lastname: Lomonte
authors.affiliation: Laboratório de Toxinologia, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro 21040-900, Brazil. anagextra@gmail.com.
authors.forename: Ana Gisele C
authors.initials: AG
authors.lastname: Neves-Ferreira
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José 11501, Costa Rica. boch31@gmail.com.
authors.forename: Andrés
authors.initials: A
authors.lastname: Feoli
authors.affiliation: Instituto de Biomedicina de Valencia, Consejo Superior de Investigaciones Científicas, Valencia 46010, Spain. jcalvete@ibv.csic.es.|Departamento de Biotecnología, Universidad Politécnica de Valencia, Valencia 46022, Spain. jcalvete@ibv.csic.es.
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Calvete
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José 11501, Costa Rica. jose.gutierrez@ucr.ac.cr.
authors.forename: José María
authors.initials: JM
authors.lastname: Gutiérrez
authors.affiliation: Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José 11501, Costa Rica. alexandra.rucavado@ucr.ac.cr.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Rucavado
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects; D002364:Caseins / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004487:Edema; D005780:Gelatin / Q000378:metabolism; D006470:Hemorrhage; D006801:Humans; D045726:Metalloproteases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D051379:Mice; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D010974:Platelet Aggregation / Q000187:drug effects; D000072417:Protein Domains; D059748:Proteolysis; D012910:Snake Venoms / Q000737:chemistry*; D017819:Viperidae; D015032:Zinc / Q000378:metabolism
chemical_list: D002364:Caseins; D018076:DNA, Complementary; D010455:Peptides; D012910:Snake Venoms; C026018:azocasein; D005780:Gelatin; D045726:Metalloproteases; D015032:Zinc
keywords: PII SVMP homologues; disintegrin domain; hemorrhagic activity; platelet aggregation; proteinase activity; snake venom metalloproteinases; zinc-binding motif