Side-chain conformational restriction in template-competitive inhibitors of E. coli DNA polymerase I Klenow fragment: synthesis, structural characterization and inhibition activity.
Side-chain conformational restriction in template-competitive inhibitors of E. coli DNA polymerase I Klenow fragment: synthesis, structural characterization and inhibition activity.
复制标题
大肠杆菌 DNA 聚合酶 I Klenow 片段模板竞争性抑制剂的侧链构象限制：合成、结构表征和抑制活性。
DOI：
10.1081/ncn-200034042
复制
发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Poolson,Tina
中科院分区：
文献类型：
--
作者：
Doughty,MichaelB;Aboudehen,Karam;Anderson,Garland;Li,Ke;Moore2nd,Bob;Poolson,Tina
以2-卤代-2-(4-叠氮苯基)乙醛为原料，分别与dATP和DAMP缩合，合成了核苷酸三磷酸α-(4-叠氮苯基)-1，N6-亚乙基-dATP3及其单磷酸3M。结构分析表明，叠氮苯基侧链连接在亚乙基的α位置(即连接在嘌呤的N1上的碳)，构象计算表明，在50°和130°的亚乙基苯键旋转中，苯环的大部分从亚乙基平面伸出，构象计算显示其最小值。与DNA Pol抑制剂2-(4-叠氮苯酰基)硫代-2‘-脱氧腺苷-5’-三磷酸1一样，核苷酸3是模板竞争性DNA聚合酶抑制剂(TCPI)，其竞争KI为3.41µM，但相对于模板竞争性逆转录酶抑制剂2-(4-叠氮苯酰基)硫代-1，N6-乙烯-2‘-脱氧腺苷5’-三磷酸2而言，其作为HIV RT抑制剂的活性较弱。此外，3以时间依赖的方式使KF失活，证实了3与游离型KF结合的动力学数据。3的TCPI活性为复合底物聚合酶抑制剂的延长侧链构象偏好提供了证据。
Nucleotide triphosphate α‐(4‐azidophenyl)‐1,N6‐etheno‐dATP3and its monophosphate3mwere synthesized by condensation of 2‐halo‐2‐(4‐azidophenyl)acetaldehyes with dATP and dAMP, respectively. Structure analysis shows that the azidophenyl side chain is attached to the α‐position of the etheno ring (i.e., the carbon attached to N1 of the purine), and conformation calculations show minima in the etheno‐phenyl bond rotation at 50 and 130° where the bulk of the phenyl ring projects out from the plane of the etheno group. Like DNA Pol inhibitor 2‐(4‐azidophenacyl)thio‐2′‐deoxyadenosine 5′‐triphosphate1, nucleotide3is a template‐competitive DNA polymerase inhibitor (TCPI), with a competitive Ki for Pol I KF of 3.41 µM, but has only weak activity as an HIV RT inhibitor relative to the template‐competitive reverse transcriptase inhibitor 2‐(4‐azidophenacyl)thio‐1,N6‐etheno‐2′‐deoxyadenosine 5′‐triphosphate2. Additionally,3photoinactivates KF in a time‐dependent manner, confirming the kinetic data that3binds to the free form of KF. The TCPI activity of3provides evidence for an extended side chain conformational preference in the combined substrate polymerase inhibitors.