The establishment and application of antiviral screening model targeted on SAH hydrolase.
- Author:
Tian ZHANG
1
;
Peizhen TAO
;
Lin WANG
;
Xin CHEN
;
Guangxing WANG
Author Information
- Publication Type:Journal Article
- MeSH: Adenine; analogs & derivatives; chemistry; pharmacology; Adenosylhomocysteinase; Animals; Antiviral Agents; chemistry; pharmacology; Drug Evaluation, Preclinical; Female; Hydrolases; antagonists & inhibitors; isolation & purification; metabolism; In Vitro Techniques; Liver; enzymology; Rats; Rats, Wistar
- From: Chinese Journal of Experimental and Clinical Virology 2002;16(1):55-58
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUNDTo establish a SAH hydrolase antiviral screening in vitro model for screening of broad spectrum antiviral agents.
METHODSSAH hydrolase was purified from rat livers by (NH4) 2SO4 fractionation, DEAE52,hydroxyapatite and Sephadex G-100 chromatography successively. The activity of SAH hydrolase was estimated by radio labeled substrate in synthesis direction by TLC.
RESULTSPurified SAH hydrolase showed a single band in SDS-PAGE electrophoresis with silver nitrate staining, the apparent molecular weight is 45 000. The Km for adenosine is (6.32 +- 0.17) micromol/L. The IC50 of S-DNPA, a known inhibitor of SAH hydrolase, was 7.6 micromol/L estimated in our system. The structure and activity relationships shown by racemic and regiosomer analogs of S-DHPA indicated that the structural specificity of SAH hydrolase was high. 42 compounds had been screened in the system and no compound showed more inhibitory activity against SAH hydrolase than S-DNPA.
CONCLUSIONSAn in vitro antiviral screening model has been established using SAH hydrolase. It can also be used to study kinetics of enzyme inhibition.
