First report of the characterization of a snake venom apyrase (Ruviapyrase) from Indian Russell's viper (Daboia russelii) venom.
Int J Biol Macromol
; 111: 639-648, 2018 May.
Article
em En
| MEDLINE
| ID: mdl-29325746
ABSTRACT
A novel apyrase from Russell's viper venom (RVV) was purified and characterized, and it was named Ruviapyrase (Russell's viper apyrase). It is a high molecular weight (79.4â¯kDa) monomeric glycoprotein that contains 2.4% neutral sugars and 58.4% N-linked oligosaccharides and strongly binds to Concanavalin A. The LC-MS/MS analysis did not identify any protein in NCBI protein database, nevertheless some de novo sequences of Ruviapyrase showed putative conserved domain of apyrase superfamily. Ruviapyrase hydrolysed adenosine triphosphate (ATP) to a significantly greater extent (pâ¯<â¯.05) as compared to adenosine diphosphate (ADP); however, it was devoid of 5'-nucleotidase and phosphodiesterase activities. The Km and Vmax values for Ruviapyrase towards ATP were 2.54⯵M and 615⯵M of Piâ¯releasedâ¯min-1, respectively with a turnover number (Kcat) of 24,600â¯min-1. Spectrofluorometric analysis demonstrated interaction of Ruviapyrase with ATP and ADP at Kd values of 0.92â¯nM and 1.25â¯nM, respectively. Ruviapyrase did not show cytotoxicity against breast cancer (MCF-7) cells and haemolytic activity, it exhibited marginal anticoagulant and strong antiplatelet activity, and dose-dependently reversed the ADP-induced platelet aggregation. The catalytic activity and platelet deaggregation property of Ruviapyrase was significantly inhibited by EDTA, DTT and IAA, and neutralized by commercial monovalent and polyvalent antivenom.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Apirase
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Venenos de Víboras
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Antivenenos
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Daboia
Limite:
Animals
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Humans
Idioma:
En
Revista:
Int J Biol Macromol
Ano de publicação:
2018
Tipo de documento:
Article