[The establishment and application of antiviral screening model targeted on SAH hydrolase].
BACKGROUND
To establish a SAH hydrolase antiviral screening in vitro model for screening of broad spectrum antiviral agents.


METHODS
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.


RESULTS
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.


CONCLUSIONS
An in vitro antiviral screening model has been established using SAH hydrolase. It can also be used to study kinetics of enzyme inhibition.
Zhang
Tian
T
Institute of Medicinal Biotechnology and Institute of Materia Medica CAMS and PUMC, Beijing 100050, China.
Tao
Peizhen
P
Wang
Lin
L
Chen
Xin
X
Wang
Guangxing
G
D000225:Adenine / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D043383:Adenosylhomocysteinase; D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical; D005260:Female; D006867:Hydrolases / Q000037:antagonists & inhibitors* / Q000302:isolation & purification / Q000378:metabolism; D066298:In Vitro Techniques; D008099:Liver / Q000201:enzymology; D051381:Rats; D017208:Rats, Wistar
D000998:Antiviral Agents; D006867:Hydrolases; D043383:Adenosylhomocysteinase; D000225:Adenine