| Isolation and functional characterization of a new acidic PLA(2) Ba SpII RP4 of the Bothrops alternatus snake venom from Argentina. | |
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MedLine Citation:
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PMID: 20331996 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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An acidic protein with phospholipase A(2) activity was purified to homogeneity from the venom of the Northeast Argentinian viperid Bothrops alternatus by two chromatographic steps: a conventional gel filtration on Sephadex G-75 and reversed phase on C18 HPLC column. A molecular mass of 14185.48 Da was determined by mass spectrometry, displaying a homodimer conformation. The kinetic assay demonstrated a catalytically active phospholipase A(2) in correspondence with Asp49 PLA(2) group. The enzyme designated Ba SpII RP4 contains an amino acid composition of 121 residues and a calculated theoretical pI value of 4.88. Amino acid sequence alignments with other Bothrops PLA(2) revealed a high degree of homology sequence (90-56%). Ba SpII RP4 did not show myotoxic activity upon muscular fibers at doses up to 100 microg i.m. route injection or lethal response when it was i.p. injected at the hightest dose of 200 microg. This toxin generates slight biological activities like paw edema inflammation and a delay in the clotting time, although Ba SpII RP4 exhibited catalytic activity. The primary amino acid sequence, determined a quadruple-time of flight (Q-TOF) hybrid mass spectrometer Q-TOF Ultima from Micromass (Manchester, UK) equipped with a nano Zspray source operating in a positive ion mode and tandem mass spectrum, an ESI/MS mass spectrum (TOF MS mode) "de novo amino acid sequencing", also provides more database about the small group of the non-myotoxic PLA(2)s isolated up to the present. |
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Authors:
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María E Garcia Denegri; Ofelia C Acosta; Salomón Huancahuire-Vega; Daniel Martins-de-Souza; Sergio Marangoni; Silvana L Maruñak; Gladys P Teibler; Laura C Leiva; Luis A Ponce-Soto |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-03-21 |
Journal Detail:
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Title: Toxicon : official journal of the International Society on Toxinology Volume: 56 ISSN: 1879-3150 ISO Abbreviation: Toxicon Publication Date: 2010 Aug |
Date Detail:
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Created Date: 2010-05-03 Completed Date: 2010-08-02 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 1307333 Medline TA: Toxicon Country: England |
Other Details:
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Languages: eng Pagination: 64-74 Citation Subset: IM |
Copyright Information:
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Copyright 2010 Elsevier Ltd. All rights reserved. |
Affiliation:
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Laboratorio de Química Biológica, Departamento de Bioquímica, Facultad de Ciencias Exactas y Naturales y Agrimensura, Universidad Nacional del Nordeste (UNNE), Av. Libertad 5470, Campus Universitario, CP 3400, Corrientes, Argentina. |
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| MeSH Terms | |
Descriptor/Qualifier:
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Alkylation Amino Acid Sequence Animals Anticoagulants* / chemistry, isolation & purification, metabolism, toxicity Argentina Bothrops* Creatine Kinase / blood Crotalid Venoms / enzymology* Edema / chemically induced Group III Phospholipases A2* / chemistry, isolation & purification, metabolism, toxicity Hemolytic Agents* / chemistry, isolation & purification, metabolism, toxicity Hydrogen-Ion Concentration Kinetics Lethal Dose 50 Mice Molecular Sequence Data Molecular Weight Muscles / drug effects, pathology Oxidation-Reduction Peptide Fragments / chemistry, isolation & purification Protein Multimerization Reptilian Proteins* / chemistry, isolation & purification, metabolism, toxicity Sequence Alignment Sequence Homology, Amino Acid |
| Chemical | |
Reg. No./Substance:
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0/Anticoagulants; 0/Crotalid Venoms; 0/Hemolytic Agents; 0/Peptide Fragments; 0/Reptilian Proteins; EC 2.7.3.2/Creatine Kinase; EC 3.1.1.4/Group III Phospholipases A2 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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