Artículos de revistas
Biochemical and biological characterization of a PLA(2) from crotoxin complex of Crotalus durissus cumanensis
Toxicon. Pergamon-elsevier Science Ltd, v. 53, n. 5, n. 534, n. 542, 2009.
A new PLA(2) (Cdcum6) from crotoxin complex of Colombian Crotalus durissus cumanensis rattlesnake was purified using molecular exclusion chromatography and RP-HPLC. The molecular mass of Cdcum6 was determined by SDS-PAGE similar to 14 KDa and confirmed by MALDI-TOF (14321.98 Da). The enzyme showed K-m 6.0 mM, V-max 3.44 nmol/min, optimum pH was 8.0 and temperature was between 30 and 45 degrees C, and it had a strict requirement of Ca2+ for its activity. The N-terminal sequence of PLA(2) was SLVQF EKMIK EVAGK NGVPWY. Comparison of amino acid sequence data with other PLA(2) from South American Crotalus durissus rattlesnakes showed that Cdcum6 shares the highest sequence identity with Cdr13 an isoform PLA(2) from Crotalus durissus ruruima, nevertheless, Cdcum6 showed high content of basic and hydrophobic amino acids. In mice, Cdcum6 presented higher LD50 than crotoxin complex from C. d. cumanensis. Additionally, Cdcum6 induced a conspicuous local myotoxic effect and moderate footpad edema; in vitro, it was antigoagulant in doses as low as 0.5 mu g/ml, and it was not cytotoxic on myoblast but Cdcum6 was able to lyse myotubes. (C) 2009 Elsevier Ltd. All rights reserved.535534542Universidad de Antioquia, COLCIENCIAS [393-2006]CYTED [206AC0281]Universidad de Antioquia, COLCIENCIAS [393-2006]CYTED [206AC0281]