Mechanism of inhibition of duck hepatitis B virus polymerase by (-)-beta-L-2',3'-dideoxy-3'-thiacytidine
Mechanism of inhibition of duck hepatitis B virus polymerase by (-)-beta-L-2',3'-dideoxy-3'-thiacytidine
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(-)-β-L-2,3-二脱氧-3-硫胞苷抑制鸭乙型肝炎病毒聚合酶的机制
DOI：
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发表时间：
1995
影响因子：
4.9
通讯作者：
D. Tyrrell
中科院分区：
文献类型：
--
作者：
Alberto Severini;XIAO;John S. Wilson;D. Tyrrell
我们利用鸭肝病毒核心颗粒的内源性逆转录酶反应来阐明核苷类似物(-)- β - l -2',3'-二脱氧-3'-硫代胞苷（3TC）抑制鸭乙型肝炎病毒（DHBV）复制的机制。与人类免疫缺陷病毒复制的情况一样，3TC-5'-三磷酸（3TC-TP）作为DNA聚合酶活性的链终止器。几个不同的实验结果支持这一结论，这解释了3TC对肝病毒的有效活性。在分离的DHBV核心颗粒中，3TC-TP抑制逆转录酶的方式类似于对dCTP的竞争性抑制。然而，在双倒数图上，最高浓度的3TC-TP和最低浓度的dCTP的抑制动力学不是线性的。如果结合到核苷酸位点之后DNA链终止，这种异常是可以预料的。仅使用曲线线性部分的计算得出Ki值为0.78 +/- 0.10微米3TC-TP。3TC-TP体外培养对核心颗粒的抑制作用不因去除游离抑制剂而逆转。3TC-TP呈浓度依赖性灭活逆转录酶活性。链终止反应的Km为0.71 +/- 0.05 μ m。对来自血清的病毒颗粒的内源性DNA聚合酶也获得了类似的竞争动力学和不可逆抑制，这表明3TC-TP也可以作为DHBV复制DNA定向DNA聚合酶的链终止器。（摘要删节250字）
We have used the endogenous reverse transcriptase reaction of viral core particles from duck liver to elucidate the mechanism of inhibition of duck hepatitis B virus (DHBV) replication by the nucleoside analog (-)-beta-L-2',3'-dideoxy-3'-thiacytidine (3TC). As is the case in human immunodeficiency virus replication, 3TC-5'-triphosphate (3TC-TP) acts as a chain terminator for the DNA polymerase activities. The results of several different experiments support this conclusion, which explains the potent activity of 3TC against the hepadnaviruses. In isolated DHBV core particles, 3TC-TP inhibited the reverse transcriptase in a manner that resembled competitive inhibition with respect to dCTP. However, the kinetics of inhibition was not linear on a double-reciprocal plot for the highest concentrations of 3TC-TP and the lowest concentration of dCTP. This anomaly would be expected if binding to the nucleotide site was followed by DNA chain termination. Calculations that used only the linear part of the curve yielded a Ki of 0.78 +/- 0.10 microM 3TC-TP. The inhibition of core particles incubated in vitro with 3TC-TP was not reversed by removal of the free inhibitor. 3TC-TP inactivated the reverse transcriptase activity in a concentration-dependent manner. The Km of the chain termination reaction was calculated at 0.71 +/- 0.05 microM. Similar competitive kinetics and irreversible inhibition were also obtained on the endogenous DNA polymerase from viral particles from serum, suggesting that 3TC-TP also acts as a chain terminator of the DNA-directed DNA polymerase of DHBV replication.(ABSTRACT TRUNCATED AT 250 WORDS)
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2,3-二脱氧-3-硫胞苷的两种顺式对映体的抗人类免疫缺陷病毒活性差异的生化基础。
DOI：
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发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Skalski,V;Chang,CN;Dutschman,G;Cheng,YC
通讯作者：
Cheng,YC
脱氧胞苷脱氨酶抗性立体异构体是 (l-)-2,3-二脱氧-3-硫胞苷在抑制乙型肝炎病毒复制中的活性形式。
DOI：
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发表时间：
1992
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Chang,CN;Doong,SL;Zhou,JH;Beach,JW;Jeong,LS;Chu,CK;Tsai,CH;Cheng,YC;Liotta,D;Schinazi,R
通讯作者：
Schinazi,R
( )- 和 (-)-2,3-二脱氧-3-硫胞苷作为抗乙型肝炎病毒剂的生化药理学。
DOI：
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发表时间：
1992
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Chang,CN;Skalski,V;Zhou,JH;Cheng,YC
通讯作者：
Cheng,YC
DOI：
10.1073/pnas.88.19.8495
发表时间：
1991-10-01
影响因子：
11.1
作者：
DOONG, SL;TSAI, CH;CHENG, YC
通讯作者：
CHENG, YC