title: Characterization of major zinc containing myonecrotic and procoagulant metalloprotease 'malabarin' from non lethal trimeresurus malabaricus snake venom with thrombin like activity: its neutralization by chelating agents.
abstract: A major myonecrotic zinc containing metalloprotease 'malabarin' with thrombin like activity was purified by the combination of gel permeation and anion exchange chromatography from T. malabaricus snake venom. MALDI-TOF analysis of malabarin indicated a molecular mass of 45.76 kDa and its N-terminal sequence was found to be Ile-Ile-Leu- Pro(Leu)-Ile-Gly-Val-Ile-Leu(Glu)-Thr-Thr. Atomic absorption spectral analysis of malabarin raveled the association of zinc metal ion. Malabarin is not lethal when injected i.p. or i.m. but causes extensive hemorrhage and degradation of muscle tissue within 24 hours. Sections of muscle tissue under light microscope revealed hemorrhage and congestion of blood vessel during initial stage followed by extensive muscle fiber necrosis with elevated levels of serum creatine kinase and lactate dehydrogenase activity. Malabarin also exhibited strong procoagulant action and its procoagulant action is due to thrombin like activity; it hydrolyzes fibrinogen to form fibrin clot. The enzyme preferentially hydrolyzes Aα followed by B subunits of fibrinogen from the N-terminal region and the released products were identified as fibrinopeptide A and fibrinopeptide B by MALDI. The myonecrotic, fibrinogenolytic and subsequent procoagulant activities of malabarin was neutralized by specific metalloprotease inhibitors such as EDTA, EGTA and 1, 10-phenanthroline but not by PMSF a specific serine protease inhibitor. Since there is no antivenom available to neutralize local toxicity caused by T. malabaricus snakebite, EDTA chelation therapy may have more clinical relevance over conventional treatment.
authors.affiliation: Department of Studies in Biochemistry, University of Mysore, Manasagangotri, Mysore-57006, Karnataka, India.
authors.forename: C D Raghavendra
authors.initials: CD
authors.lastname: Gowda
authors.forename: H V
authors.initials: HV
authors.lastname: Shivaprasad
authors.forename: R Venkatesh
authors.initials: RV
authors.lastname: Kumar
authors.forename: R
authors.initials: R
authors.lastname: Rajesh
authors.forename: Y K
authors.initials: YK
authors.lastname: Saikumari
authors.forename: B M
authors.initials: BM
authors.lastname: Frey
authors.forename: F J
authors.initials: FJ
authors.lastname: Frey
authors.forename: B K
authors.initials: BK
authors.lastname: Sharath
authors.forename: B S
authors.initials: BS
authors.lastname: Vishwanath
mesh_terms: D000818:Animals; D000997:Antivenins / Q000737:chemistry / Q000494:pharmacology; D001777:Blood Coagulation; D002614:Chelating Agents / Q000737:chemistry / Q000494:pharmacology*; D002850:Chromatography, Gel; D003402:Creatine Kinase / Q000032:analysis / Q000378:metabolism; D003435:Crotalid Venoms / Q000037:antagonists & inhibitors* / Q000201:enzymology / Q000633:toxicity; D004492:Edetic Acid / Q000737:chemistry / Q000494:pharmacology*; D006470:Hemorrhage / Q000188:drug therapy* / Q000473:pathology / Q000517:prevention & control; D007770:L-Lactate Dehydrogenase / Q000032:analysis / Q000378:metabolism; D008297:Male; D045726:Metalloproteases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000302:isolation & purification / Q000633:toxicity; D051379:Mice; D008970:Molecular Weight; D009132:Muscles / Q000187:drug effects / Q000473:pathology; D009336:Necrosis / Q000188:drug therapy* / Q000473:pathology / Q000517:prevention & control; D012909:Snake Bites; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D017841:Trimeresurus / Q000502:physiology*; D015032:Zinc / Q000378:metabolism
chemical_list: D000997:Antivenins; D002614:Chelating Agents; D003435:Crotalid Venoms; D004492:Edetic Acid; D007770:L-Lactate Dehydrogenase; D003402:Creatine Kinase; D045726:Metalloproteases; D015032:Zinc