The establishment and application of antiviral screening model targeted on SAH hydrolase / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>BACKGROUND</b>To establish a SAH hydrolase antiviral screening in vitro model for screening of broad spectrum antiviral agents.</p><p><b>METHODS</b>SAH 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.</p><p><b>RESULTS</b>Purified 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.</p><p><b>CONCLUSIONS</b>An in vitro antiviral screening model has been established using SAH hydrolase. It can also be used to study kinetics of enzyme inhibition.</p>