Artículos de revistas
Biological and biochemical characterization of two new PLA(2) isoforms Cdc-9 and Cdc-10 from Crotalus durissus cumanensis snake venom
Registro en:
Comparative Biochemistry And Physiology C-toxicology & Pharmacology. Elsevier Science Inc, v. 151, n. 1, n. 66, n. 74, 2010.
1532-0456
WOS:000273152400008
10.1016/j.cbpc.2009.08.011
Autor
Romero-Vargas, FF
Ponce-Soto, LA
Martins-de-Souza, D
Marangoni, S
Institución
Resumen
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) This work reports the purification, biological characterization and amino acid sequence of two new basic PLA(2) isoforms, Cdc-9 and Cdc-10, purified from the Crotalus durissus cumanensis venom by one step analytical chromatography reverse phase HPLC. The molecular masses of the PLA(2) were 14,175 +/- 2.7 Da for Cdc-9 and 14,228 +/- 3.5 Da for Cdc-10 both deduced by primary structure and confirmed by MALDI-TOF. The isoforms presented an amino acid sequence of 122 amino acid residues, being Cdc-9: SLVQFNKMIK FETRKSGLPF YAAYGCYCGW GGQRPKDATD RCCFVHDCCY GKVAKCNTKW DIYSYSLKSG YITCGKGTWC KEQICECDRV AAECLRRSLS TYKNEYMFYP DSRCREPPEY TC with pI value of 8.25 and Cdc-10: SLLQFNKMIK FETRKSGVPF YAAYGCYCGW GGRRPKDPTD RCCFVHDCCY GKLTKCNTKW DIYSYSLKSG YITCGKGTWC KEQICECDRV AAECLRRSLN TYKNEYMFYP DSRCRGPPEY TC with a pI value of 8.46, showing highly conserved Ca2+-binding and catalytic sites. The PLA(2) activity decreased when the isoforms Cdc-9 and Cdc-10 were incubated with 4-bromophenacyl bromide (p-BPB), anhydrous acetic acid and p-nitrobenzene sulfonyl fluoride (NBSF) when compared with the activity of both native isoforms. In mice, the PLA(2) isoforms Cdc-9 and Cdc-10 induced myonecrosis and edema. Myotoxic and edema activities were reduced after treatment of the isoforms with p-BPB; acetylation of the lysine residues and the treatment of PLA(2) with NBSF have also induced edema reduction. However, p-BPB strongly diminishes the local and systemic myotoxic effects. (C) 2009 Elsevier Inc. All rights reserved. 151 1 66 74 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)