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Purification and activity of two phospholipase enzymes from Naja nigricolis nigricolis reinhardt venom
Author(s) -
Abubakar M. S.,
Nok A. J.,
Abdurahman E. M.,
Haruna A. K.,
Shok M.
Publication year - 2003
Publication title -
journal of biochemical and molecular toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.526
H-Index - 58
eISSN - 1099-0461
pISSN - 1095-6670
DOI - 10.1002/jbt.10060
Subject(s) - sephadex , chemistry , enzyme , size exclusion chromatography , chromatography , venom , biochemistry , hydrolysis , monomer , phospholipase , phospholipase a , phospholipase a2 , organic chemistry , polymer
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