Enzymatic analysis of Hemiscorpius lepturus scorpion venom using zymography and venom-specific antivenin.
Hemiscorpius lepturus envenomation exhibits various pathological changes in the affected tissues, including skin, blood cells, cardiovascular and central nervous systems. The enzymatic activity and protein component of the venom have not been described previously. In the present study, the electrophoretic profile of H. lepturus venom was determined by SDS-PAGE (12 and 15%), resulting in major protein bands at 3.5-5, 30-35 and 50-60 kDa. The enzymatic activities of the venom was, for the first time, investigated using various zymography techniques, which showed the gelatinolytic, caseinolytic, and hyaluronidase activities mainly at around 50-60 kDa, 30-40 kDa, and 40-50 kDa, respectively. Among these, the proteolytic activities was almost completely disappeared in the presence of a matrix metalloproteinase inhibitor, 1, 10-phenanthroline. Antigen-antibody interactions between the venom and its Iranian antivenin was observed by Western blotting, and it showed several antigenic proteins in the range of 30-160 kDa. This strong antigen-antibody reaction was also demonstrated through an enzyme-linked immunosorbent assay (ELISA). The gelatinase activity of the venom was suppressed by Razi institute polyvalent antivenin, suggesting the inhibitory effect of the antivenin against H. lepturus venom protease activities. Prudently, more extensive clinical studies are necessary for validation of its use in envenomed patients.
Seyedian
Ramin
R
Department of Pharmacology and Toxicology, Jundishapur University of Medical Sciences, Ahvaz, Iran.
Pipelzadeh
Mohammad Hassan
MH
Jalali
Amir
A
Kim
Euikyung
E
Lee
Hyunkyoung
H
Kang
Changkeun
C
Cha
Mijin
M
Sohn
Eun-Tae
ET
Jung
Eun-Sun
ES
Rahmani
Ali Hassan
AH
Mirakabady
Abbas Zare
AZ
D000818:Animals; D000997:Antivenins / Q000737:chemistry* / Q000494:pharmacology; D015153:Blotting, Western; D002364:Caseins / Q000737:chemistry; D002417:Cattle; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005780:Gelatin / Q000737:chemistry; D018093:Gelatinases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification; D006820:Hyaluronic Acid / Q000737:chemistry; D006821:Hyaluronoglucosaminidase / Q000737:chemistry / Q000302:isolation & purification; D008666:Metalloendopeptidases / Q000737:chemistry / Q000302:isolation & purification; D012604:Scorpion Venoms / Q000737:chemistry / Q000201:enzymology* / Q000494:pharmacology; D012605:Scorpions / Q000201:enzymology*; D013552:Swine
D000997:Antivenins; D002364:Caseins; D004791:Enzyme Inhibitors; D012604:Scorpion Venoms; D005780:Gelatin; D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase; D018093:Gelatinases; D008666:Metalloendopeptidases; C030895:caseinase