SYNTHESIS AND BIOCHEMICAL EVALUATION OF 2'-DEOXY-LIN-BENZOADENOSINE PHOSPHATES
SYNTHESIS AND BIOCHEMICAL EVALUATION OF 2'-DEOXY-LIN-BENZOADENOSINE PHOSPHATES
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DOI：
10.1021/bi00312a012
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发表时间：
1984-01-01
期刊：
影响因子：
2.9
通讯作者：
LEONARD, NJ
中科院分区：
文献类型：
--
作者：
LESSOR, RA;GIBSON, KJ;LEONARD, NJ
2“-脱氧-lin-苯并腺苷是通过2-（β-吲哚基）腺苷的还原脱氧合成的。D-呋喃核糖基）-8-（甲硫基）咪唑并[4，5-g]喹唑啉。通过化学和/或酶促方法制备5“-单磷酸、5”-二磷酸和5“-三磷酸。5 ''-二磷酸是[兔肌肉]丙酮酸激酶磷酸化的底物，并在动力学测定中与各种天然和扩展底物进行了比较。当在用大肠杆菌DNA聚合酶I的切口平移实验中测试2“-脱氧-lin-苯并腺苷5”-三磷酸时，从[α-β-腺苷酸]掺入的32 P水平非常低。观察到~（32）P]TTP进入poly[d（AT）]。最近邻分析表明，类似物没有显着纳入聚合物的内部位置。在DNA测序反应中，类似物在腺嘌呤残基处引起链终止，尽管终止不如2“，3”-双脱氧-ATP均匀且效率低。lin-Benzoadenine可以与胸腺嘧啶形成加宽的Watson-Crick碱基对。酶可以能够将类似物连接到DNA。
2''-Deoxy-lin-benzoadenosine was synthesized via reductive deoxygenation of 2-(.beta.-D-ribofuranosyl)-8-(methylthio)imidazo[4,5-g]quinazoline. The 5''-mono-, 5''-di- and 5''-triphosphates were prepared by chemical and/or enzymatic methods. The 5''-diphosphate was a substrate for phosphorylation by [rabbit muscle] pyruvate kinase and was compared with various natural and extended substrates in kinetic assays. When 2''-deoxy-lin-benzoadenosine 5''-triphosphate was tested in a nick-translation experiment with Escherichia coli DNA polymerase I, a very low level of 32P incorporation from [.alpha.-32P]TTP into poly[d(AT)] was observed. Nearest-neighbor analysis indicated that the analog was not significantly incorporated into internal positions in the polymer. In DNA-sequenching reactions, the analog caused chain termination at adenine residues, although termination was less uniform and less efficient than that with 2'',3''-dideoxy-ATP. lin-Benzoadenine can form a widened Watson-Crick base pair with thymine. The enzyme may be able to attach the analog to DNA.