Biological and biochemical characterization of two new PLA2 isoforms Cdc-9 and Cdc-10 from Crotalus durissus cumanensis snake venom

Frey Francisco Romero Vargas, Luis Alberto Ponce-Soto, Daniel Martins-de-Souza, Sergio Marangoni

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

This work reports the purification, biological characterization and amino acid sequence of two new basic PLA2 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 PLA2 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 PLA2 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 PLA2 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 PLA2 with NBSF have also induced edema reduction. However, p-BPB strongly diminishes the local and systemic myotoxic effects.

Original languageEnglish
Pages (from-to)66-74
Number of pages9
JournalComparative Biochemistry and Physiology - C Toxicology and Pharmacology
Volume151
Issue number1
DOIs
StatePublished - Jan 2010
Externally publishedYes

Bibliographical note

Funding Information:
The authors thank Marino Forner for general technical help. This work was supported by CAPES and is part of Ms Sc thesis by Frey Francisco Romero-Vargas.

Keywords

  • Crotalus durissus cumanensis
  • Myotoxins
  • RP-HPLC
  • Snake venom

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