Isolation and characterization of the thrombin-like enzyme from Cryptelytrops albolabris (white-lipped tree viper) venom

Tan, N.H. and Fung, S.Y. and Yap, Y.H.Y. (2012) Isolation and characterization of the thrombin-like enzyme from Cryptelytrops albolabris (white-lipped tree viper) venom. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 161 (1). pp. 79-85. ISSN 1096-4959,

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Abstract

A thrombin-like enzyme (termed albolabrase) was isolated in purified form from the venom of Cryptelytrops albolabris (white-lipped tree viper) using high performance anion ion exchange and gel filtration chromatography. The molecular mass of albolabrase was 33.7 kDa as determined by SDS-PAGE and 35.8 kDa as determined by Superose gel filtration chromatography. The N-terminal sequence was determined to be VVGGDECNINE which is homologous to many snake venom thrombin-like enzymes. Albolabrase exhibits both arginine ester hydrolase and arginine amidase activities and the enzyme is fastidious towards tripeptide chromogenic anilide substrates. The fibrinogen clotting activity was optimum at 3 mg/mL bovine fibrinogen, and showed distinct species differences in the following decreasing order: bovine fibrinogen > dog fibrinogen � human fibrinogen > goat fibrinogen. The enzyme failed to clot both rabbit and cat fibrinogens. Reversed-phase HPLC analysis on the breakdown products of fibrinogenolytic action of albolabrase indicated that the enzyme belongs to the AB class of snake venom thrombin-like enzyme. In the indirect ELISA, IgG anti-albolabrase reacted extensively with most crotalid venoms, except with Tropidolaemus wagleri and Calloselasma rhodostoma venoms. The double sandwich ELISA, however, showed that anti-albolabrase reacted strongly only with venoms from the Trimeresurus complex, and that the results support the proposed new taxonomy changes concerning the Trimeresurus complex.

Item Type:	Article
Funders:	UNSPECIFIED
Additional Information:	Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia ---> Corresponding author. Tel.: +60 3 79674912; fax: +60 3 79674957. E-mail address: tanngethong@yahoo.com.sg (N.H. Tan).
Uncontrolled Keywords:	Cryptelytrops albolabris venom; Thrombin-like enzyme; ELISA
Subjects:	R Medicine
Divisions:	Faculty of Medicine
Depositing User:	Fung Shin Yee
Date Deposited:	25 Sep 2012 02:25
Last Modified:	25 Sep 2012 02:25
URI:	http://eprints.um.edu.my/id/eprint/3696

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