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

Raghavendra Gowda, H. V. Shivaprasad, R. Venkatesh Kumar, R. Rajesh, Y. K. Saikumari, B. M. Frey, F. J. Frey, B. K. Sharath, B. S. Vishwanath

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

A major myonecrotic zinc containing metalloprotease 'malabarin' with thrombin like activity was purified bythe combination of gel permeation and anion exchange chromatography from T. malabaricus snake venom. MALDI-TOFanalysis 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 ofzinc metalion. 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 Bβ 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 specificserine 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.

Original languageEnglish (US)
Pages (from-to)2578-2588
Number of pages11
JournalCurrent Topics in Medicinal Chemistry
Volume11
Issue number20
StatePublished - Oct 1 2011

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Trimeresurus
Snake Venoms
Metalloproteases
Chelating Agents
Thrombin
Zinc
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Edetic Acid
Muscles
Fibrinogen
Fibrinopeptide B
Fibrinopeptide A
Hemorrhage
Chelation Therapy
Antivenins
Snake Bites
Egtazic Acid
Creatine Kinase
Protease Inhibitors
Fibrin

All Science Journal Classification (ASJC) codes

  • Drug Discovery

Cite this

Gowda, Raghavendra ; Shivaprasad, H. V. ; Venkatesh Kumar, R. ; Rajesh, R. ; Saikumari, Y. K. ; Frey, B. M. ; Frey, F. J. ; Sharath, B. K. ; Vishwanath, B. S. / 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. In: Current Topics in Medicinal Chemistry. 2011 ; Vol. 11, No. 20. pp. 2578-2588.
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abstract = "A major myonecrotic zinc containing metalloprotease 'malabarin' with thrombin like activity was purified bythe combination of gel permeation and anion exchange chromatography from T. malabaricus snake venom. MALDI-TOFanalysis 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 ofzinc metalion. 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 Bβ 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 specificserine 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.",
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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. / Gowda, Raghavendra; Shivaprasad, H. V.; Venkatesh Kumar, R.; Rajesh, R.; Saikumari, Y. K.; Frey, B. M.; Frey, F. J.; Sharath, B. K.; Vishwanath, B. S.

In: Current Topics in Medicinal Chemistry, Vol. 11, No. 20, 01.10.2011, p. 2578-2588.

Research output: Contribution to journalArticle

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T1 - Characterization of major zinc containing myonecrotic and procoagulant metalloprotease 'malabarin' from non lethal Trimeresurus malabaricus snake venom with thrombin like activity

T2 - Its neutralization by chelating agents

AU - Gowda, Raghavendra

AU - Shivaprasad, H. V.

AU - Venkatesh Kumar, R.

AU - Rajesh, R.

AU - Saikumari, Y. K.

AU - Frey, B. M.

AU - Frey, F. J.

AU - Sharath, B. K.

AU - Vishwanath, B. S.

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N2 - A major myonecrotic zinc containing metalloprotease 'malabarin' with thrombin like activity was purified bythe combination of gel permeation and anion exchange chromatography from T. malabaricus snake venom. MALDI-TOFanalysis 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 ofzinc metalion. 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 Bβ 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 specificserine 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.

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