Analysis of phospholipase A2, L-amino acid oxidase, and proteinase enzymatic activities of the Lachesis muta rhombeata venom.
The study of venom components is an important step toward understanding the mechanism of action of such venoms and is indispensable for the development of new therapies. This work aimed to investigate the venom of Lachesis muta rhombeata and evaluate enzymes related to its toxicity. Phospholipase A2 (PLA(2)), L-amino acid oxidase (LAAO), and proteinase activities were measured, and the molecular weights were estimated. We found the venom to contain one PLA(2) (17 kDa), one LAAO (132 kDa), and three serine proteinases (40, 31, and 20 kDa). Although only serine proteinases were observed in the zymogram, metalloproteinases were found to contribute more to the total proteolytic activity than did serine proteinases. The work confirmed the presence of highly active enzymes; and, moreover, we proposed a novel method for confirming the presence of LAAOs by zymography. We also suggested a simple step to increase the sensitivity of proteinase assays.
Department of Biochemistry and Immunology, University of São Paulo at Ribeirão Preto School of Medicine 14049-900, Ribeirão Preto, Brazil.
Lucas Benício
LB
Campos
Manuela Berto
MB
Pucca
Eduardo Crosara
EC
Roncolato
Joaquim Coutinho
JC
Netto
José Elpidio
JE
Barbosa
D000818:Animals; D002364:Caseins / Q000737:chemistry; D002642:Chick Embryo; D004912:Erythrocytes / Q000187:drug effects; D006461:Hemolysis / Q000187:drug effects; D050830:L-Amino Acid Oxidase / Q000737:chemistry*; D007930:Leucine / Q000737:chemistry; D051379:Mice; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010447:Peptide Hydrolases / Q000737:chemistry*; D054467:Phospholipases A2 / Q000737:chemistry* / Q000494:pharmacology; D011480:Protease Inhibitors / Q000737:chemistry; D059748:Proteolysis; D012710:Serum Albumin, Bovine / Q000737:chemistry; D014757:Viper Venoms / Q000737:chemistry / Q000201:enzymology* / Q000494:pharmacology; D017819:Viperidae
D002364:Caseins; D011480:Protease Inhibitors; D014757:Viper Venoms; D012710:Serum Albumin, Bovine; D050830:L-Amino Acid Oxidase; D054467:Phospholipases A2; D010447:Peptide Hydrolases; D007930:Leucine