Purification, characterization and gene cloning of a novel phospholipase A2 from the venom of Agkistrodon blomhoffii ussurensis

Int J Biochem Cell Biol. 2005 Mar;37(3):558-65. doi: 10.1016/j.biocel.2004.05.022.

Abstract

A new phospholipase A2 with Gln at the site 49, abbreviated as Gln49-PLA2, has been purified from the venom of Agkistrodon blomhoffii ussurensis by using ion-exchange chromatography, gel filtration chromatography and reversed-phase HPLC, and behaves as a single-band on SDS-PAGE. Its molecular weight is 13881.85+/-0.33 Da given by mass spectrometry and pI is about 8.56 given by isoelectric focusing. Gln49-PLA2 does not show phospholipase A2 and hemorrhagic activity, whereas shows weak toxic and apparent anticoagulant activity. Based on the N-terminal sequencing and peptide mass fingerprint analysis, Gln49-PLA2 cDNA has been cloned by means of RT-PCR. Gln49-PLA2 consists of 122 amino acid residues and has the structural features of class II of snake venom phospholipase A2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agkistrodon*
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Chromatography, High Pressure Liquid
  • Chromatography, Ion Exchange
  • Cloning, Molecular*
  • Crotalid Venoms / genetics*
  • Crotalid Venoms / isolation & purification*
  • Electrophoresis, Polyacrylamide Gel
  • Glycine / chemistry
  • Isoelectric Focusing
  • Mass Spectrometry
  • Molecular Sequence Data
  • Molecular Weight
  • Peptide Mapping
  • Phospholipases A / chemistry
  • Phospholipases A / genetics*
  • Phospholipases A / isolation & purification*
  • Phospholipases A2
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, Protein

Substances

  • Crotalid Venoms
  • Phospholipases A
  • Phospholipases A2
  • Glycine