Presteady state kinetic investigation of the incorporation of anti-hepatitis B nucleotide analogues catalyzed by noncanonical human DNA polymerases.
Presteady state kinetic investigation of the incorporation of anti-hepatitis B nucleotide analogues catalyzed by noncanonical human DNA polymerases.
复制标题
抗肝炎B核苷酸类似物的掺入型核苷酸类似物的质疑状态动力学研究。
DOI：
10.1021/tx200458s
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发表时间：
2012-01-13
影响因子：
4.1
通讯作者：
Suo Z
中科院分区：
文献类型：
--
作者：
Brown JA;Pack LR;Fowler JD;Suo Z
抗病毒核苷类似物已被开发用于抑制乙型肝炎病毒（HBV）聚合酶的酶活性，从而阻止HBV的复制和产生。然而，这些类似物的使用可能受到药物毒性的限制，因为这些核苷类似物（核苷酸类似物）的5 ' -三磷酸是人类DNA聚合酶整合到宿主DNA中的潜在底物。虽然它们是人类复制性DNA聚合酶的不良底物，但这些核苷酸类似物是否为最近发现的人类X和y家族DNA聚合酶的底物仍有待确定。利用稳态前动力学技术，我们测量了人类DNA聚合酶β、λ、η、ι、κ和Rev1的底物特异性值，这些酶结合了以下抗hbv核苷类似物的活性形式，这些类似物被批准用于临床：阿德福韦、替诺福韦、拉米夫定、替比夫定和恩替卡韦。与天然核苷酸的掺入相比，大多数核苷酸类似物被六种人类DNA聚合酶掺入的效率较低（2至122,000）。此外，恩替卡韦和替比夫定这两种具有3 ' -羟基的药物，通过引物延伸和DNA连接试验检测了嵌入人类DNA的潜力。这些结果表明替比夫定作为链终止物，而恩替卡韦被这六种酶有效地延长，并且是人类DNA连接酶i的底物。我们的研究结果表明，由人类X和y家族聚合酶催化的抗hbv核苷酸类似物的掺入可能有助于临床毒性。
Antiviral nucleoside analogs have been developed to inhibit the enzymatic activities of the hepatitis B virus (HBV) polymerase, thereby preventing the replication and production of HBV. However, the usage of these analogs can be limited by drug toxicity because the 5′-triphosphates of these nucleoside analogs (nucleotide analogs) are potential substrates for human DNA polymerases to incorporate into host DNA. Although they are poor substrates for human replicative DNA polymerases, it remains to be established whether these nucleotide analogs are substrates for the recently discovered human X- and Y-family DNA polymerases. Using pre-steady state kinetic techniques, we have measured the substrate specificity values for human DNA polymerases β, λ, η, ι, κ, and Rev1 incorporating the active forms of the following anti-HBV nucleoside analogs approved for clinical use: adefovir, tenofovir, lamivudine, telbivudine, and entecavir. Compared to the incorporation of a natural nucleotide, most of the nucleotide analogs were incorporated less efficiently (2 to >122,000) by the six human DNA polymerases. In addition, the potential for entecavir and telbivudine, two drugs which possess a 3′-hydroxyl, to become embedded into human DNA was examined by primer extension and DNA ligation assays. These results suggested that telbivudine functions as a chain terminator while entecavir was efficiently extended by the six enzymes and was a substrate for human DNA ligase I. Our findings suggested that incorporation of anti-HBV nucleotide analogs catalyzed by human X- and Y-family polymerases may contribute to clinical toxicity.
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影响因子：
4.8
作者：
Fowler, Jason D.;Brown, Jessica A.;Suo, Zucai
通讯作者：
Suo, Zucai
影响因子：
4.9
作者：
Feng, JY;Murakami, E;Anderson, KS
通讯作者：
Anderson, KS
影响因子：
4.8
作者：
Fiala, Kevin A.;Duym, Wade W.;Suo, Zucai
通讯作者：
Suo, Zucai
影响因子：
5.6
作者：
Brown, Jessica A.;Duym, Wade W.;Suo, Zucai
通讯作者：
Suo, Zucai
影响因子：
4.9
作者：
Birkus, G;Hájek, M;Otová, B
通讯作者：
Otová, B