Purification and activity of two phospholipase enzymes from Naja nigricolis nigricolis reinhardt venom

✍ Scribed by M. S. Abubakar; A. J. Nok; E. M. Abdurahman; A. K. Haruna; M. Shok

• Publisher: John Wiley and Sons
• Year: 2003
• Tongue: English
• Weight: 185 KB
• Volume: 17
• Category: Article
• ISSN: 1095-6670
• DOI: 10.1002/jbt.10060

✦ Synopsis

Abstract

Two phospholipase enzymes NN1 and NN2 were purified from the venom of Naja nigricolis nigricolis Reinhardt to apparent homogeneity. NN1 was purified by a two‐step anion‐exchange chromatography on DEAE‐cellulose column while NN2 was purified by a combination of anion‐exchange chromatography and gel filteration on Sephadex G‐150. The enzyme NN1 moved homogenously on acrylamide gel as a monomer with a molecular weight of 65 kDa while NN2 was a dimer of 71 kDa. Both enzymes were clearly separated. Both enzymes hydrolyzed L‐α‐phosphatidyl choline with activities of 345.5 for NN1 and 727.8 μmol min^−1^ mg^−1^ for NN2. The dimeric 71‐kDa enzyme has a higher haemolytic and anticoagulant activity than the monomeric 65‐kDa enzyme. It is apparent that the dimeric enzyme has a more pronounced activity than the monomer has, thus toxic activity may be related to the hydrolysis of phospholipids. © 2003 Wiley Periodicals, Inc. J Biochem Mol Toxicol 17:53–58, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.10060