I= CHARACTERIZATION OF VENOM PROTEINS FROM PSEUDOCERASTES PERSICUS PERSICUS
Pakistan Research Repository Home
 

Title of Thesis
CHARACTERIZATION OF VENOM PROTEINS FROM PSEUDOCERASTES PERSICUS PERSICUS

Author(s)
Farzaneh Mirzajiani
Institute/University/Department Details
University of Karachi/ H.E.J. Research Institute of Chemistry
Session
1999
Subject
Chemistry
Number of Pages
110
Keywords (Extracted from title, table of contents and abstract of thesis)
venom, proteins, false horn viper, snake, psuedocerastes persicus persicus, enzymes, haemorrhagic proteins, proteolytic proteins, anticoagulant proteins, cytotoxic proteins, enzymatic proteins, homodimer protein

Abstract
Psuedocerastes persicus persicus (false horn viper) snake venom and its gel filtration fractions were tested for a number of biological tests. Results from biological studies indicated that venom from Psuedocerastes persicus persicus contains number of toxic, haemorrhagic, proteolytic, anticoagulant, cytotoxic and enzymatic proteins. Venom did not show either haemolytic or apoptotic activity and also there was no evidence of presence of growth factors in crude venom. Among all enzymes activities tests in curde venom i.e LDH, ALT, SOD, Catalase, ALP, CPK, AST, a-Amylase, PDE, Endonuclease and L-amino acid oxidase actgivity, very high activity of LAO and moderate activity of LDH, ALP, and a-Amylase was detected in crude venom

The major lethal factor showing strong L-amino acid oxidase activity isolated from the venom of Psuedocerastes persicus persicus by a combination of gel filtration on Sephadex G-50, Sephadex G-100 and heparin affinity chromatography, is a homodimer protein with approximate mol. Wt. 13.000 Da as determined by SDS-? AGE. It is the main portion of total venom protein and exhibits potent lethal, heamorrhagic and cytotoxic activity

The enzyme exhibited unusual thermal stability and unlike most other venom LAO’s it was stable in alkaline solution and was inactivated by freezing. Kinetics studies proved that this enzyme is new isoform of L-amino acid oxidase

Cytotoxicity effect of venom and its fractions on many human and mouse cancerous cell lines including SK-CO-1, HT144, HI299, HT157, EBV-tranformed B-lymphocytes, SP2, and normal B-lymphocytes, shows that the extent of cytotoxicity effect of venom compounds is variable depending on cell lines. Cytotoxicity of crude venom and fraction (F1) containing L-amino acid oxidase was mostly inhibited by catalase, indicating that H2O2 produced by LAO is the main cause pf sutptpxocoty by venom. These data suggests the treatment with anti-oxidants for protecting tissues affected by snake bite

No apoptotic effect was detected in a number of experiments carried out on human cancerous cells (SK-CO-1 and HT144) and B-lymphocytes exposed to the different concentration of crude venom and F1 (L-amino acid oxidase), indicating that LAO is capable of inducing apoptosis in some particular cell lines but not in all cell lines

Download Full Thesis
1208.03 KB
S. No. Chapter Title of the Chapters Page Size (KB)
1 0 Contents
102.84 KB
2 1 Introduction 1
245.69 KB
  1.1 Composition of snake venoms; Their medical and scientific uses 2
  1.2 Nonproteinus snake venom compounts 15
  1.3 Description of Pseudocerastes persicus persicus 17
  1.4 Objective of the present study 18
3 2 Experimental 19
233.74 KB
  2.1 SDS-PAGE Electrophoresis 20
  2.2 Gel permeation HPLC 20
  2.3 Gel Filtration on Sephadex G-50 21
  2.4 Biological activity tests 21
  2.5 Studies on L-Amino acid Oxidase 35
4 3 Results 40
438.94 KB
  3.1 SDS-Polyacrylamide gel electrophoresis 40
  3.2 Gel permeation chromatography cf crude venom on HPLC column 40
  3.3 Gel filtration chromatography of crude venom on sephadex G-50 column 40
  3.4 Biological activity test 40
  3.5 Studies on L-Amino Acid Oxidase 42
5 4 Discussion 81
123.45 KB
6 5 Conclusion 92
213.27 KB
  5.1 References 94