Effect and mechanism of beta-L-D4A on DNA polymerase alpha.
OBJECTIVE
To investigate the safety of beta-L-D4A on DNA polymerase alpha.


METHODS
Ion exchange chromatography was used to separate DNA polymerase alpha from crude extract of human Hela cells. Detailed kinetic parameters were determined for beta-L-D4A against DNA polymerase alpha.


RESULTS
DNA polymerase alpha was purified with 4% yield and 31000 units/mg specific activity. The Michaelis constant (Km = 3.22 micromol/L), 50% inhibition concentration (IC50 = 178.49 micromol/L) and inhibition constant (Ki = 126 micromol/L) of beta-L-D4A were determined by kinetic analysis.


CONCLUSIONS
beta-L-D4A is a more safe nucleoside for hepatitis B virus infection with a lower host toxicity.
Li
Yan
Y
Institute of Liver Diseases, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Street, Wuhan 430030, Hubei Province, China.
Lin
Ju-Sheng
JS
Zhang
Ying-Hui
YH
Wang
Xiao-Yan
XY
Chang
Ying
Y
He
Xing-Xing
XX
D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D004256:DNA Polymerase I / Q000187:drug effects*; D016048:Dideoxyadenosine / Q000009:adverse effects / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D006367:HeLa Cells; D006509:Hepatitis B / Q000188:drug therapy; D006515:Hepatitis B virus / Q000502:physiology; D006801:Humans; D009705:Nucleosides / Q000009:adverse effects / Q000494:pharmacology* / Q000627:therapeutic use; D014779:Virus Replication / Q000187:drug effects
D009705:Nucleosides; D016048:Dideoxyadenosine; C075923:2'3'-didehydro-2'3'-dideoxyadenosine; D004256:DNA Polymerase I