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

Tan, N.H.; Fung, S.Y.; 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
    Creators:
    1. Tan, N.H.
    2. Fung, S.Y.(Department of Molecular Medicine, Faculty of Medicine Building, University of Malaya, 50603 Kuala Lumpur, MALAYSIA)
    3. Yap, Y.H.Y.
    Journal or Publication Title: Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology
    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 10:25
    Last Modified: 25 Sep 2012 10:25
    URI: http://eprints.um.edu.my/id/eprint/3696

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