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Effect of β-L-D4A on human DNA polymerases β and δ and its mechanism

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Author:: No author available

Journal Title:: WORLD CHINESE JOURNAL OF DIGESTOLOGY

Issue:: 6

DOI:: 10.3969/j.issn.1009-3079.2008.06.009

Key Word:: 核苷类;β-L-D4A;人DNA聚合酶β和δ;酶动力;毒副作用

Abstract：目的:对人DNA聚合酶δ进行纯化和鉴定,通过检测β-L-D4A对人DNA聚合酶β和δ的作用,观察β-L-D4A的毒副作用.方法:运用多次离子交换层析的方法提取、纯化人Hela细胞的DNA聚合酶δ:检测β-L-D4A对人DNA聚合酶β和δ的酶动力学作用.结果:提纯并鉴定了DNA聚合酶δ,收得率是15%,比活力是1911 Ukat/mg.β-L-D4A为抑制剂时,对人DNA聚合酶β和δ的酶动力学参数分别是Km=1.86 μmol/L,2.45μmol/L,IC50=25.21 μmol/L和150.1μmol/L,Ki=24.03μmol/L和132.7μmol/L,结果均提示β-L-D4A对DNA聚合酶β和δ的抑制作用远小于拉米夫定,结论:β-L-D4A是人DNA聚合酶β和δ的竞争性抑制剂,他对这两种酶的毒副作用远小于拉米夫定,可望成为更高效低毒的抗HBV类药物.

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