Direct synthesis and identification of benzo[a]pyrene diol epoxide-deoxyguanosine binding sites in modified oligodeoxynucleotides.
Direct synthesis and identification of benzo[a]pyrene diol epoxide-deoxyguanosine binding sites in modified oligodeoxynucleotides.
复制标题
修饰寡脱氧核苷酸中苯并[a]芘二醇环氧化物-脱氧鸟苷结合位点的直接合成和鉴定。
DOI：
10.1021/tx00030a007
复制
发表时间：
1992
影响因子：
4.1
通讯作者：
Geacintov,NE
中科院分区：
文献类型：
--
作者：
Mao,B;Margulis,LA;Li,B;Ibanez,V;Lee,H;Harvey,RG;Geacintov,NE
苯并[a]芘代谢物模型化合物（+）-ANFI-7反应生成的加合物|8，8 α-二羟基-9 α，10 α-环氧基-7，8，9，10-四氢苯并[a]芘[（+）-BPDE]与单链寡脱氧核苷酸5 ′-d（TATGCGTAT）根据先前描述的直接合成技术获得[Cosman，M.，Ibanez，V.，Geacintov，N. E、和Harvey，R. G.（1990）Carcinogenesis 11，1667-1672]。（+）-BPDE与鸟嘌呤G4和Ge的环外氨基（数字表示从5 '侧开始计数的鸟嘌呤位置）的反式和顺式加成（反式/顺式加成比 * 4.5）得到了四种主要的加合物。这些加合物可以通过反相高效液相色谱法彼此分离。BPDE结合在64或G n上的位点可以从经历G+ A和G Maxam-Gilbert链切割反应的BPDE修饰的寡核苷酸的20%聚丙烯酰胺凝胶上的电泳带型来确定[Maxam，A. M.，和吉尔伯特，W.（1980）方法。Enzymol. 65，499-560]。未修饰的DNA和两种不同的BPDE修饰的寡核苷酸的电泳凝胶带型不同，因为（1）带有BPDE残基的链切割片段比来自未修饰的寡核苷酸的相应片段迁移得慢，（2）在G+ A反应中，在BPDE修饰的鸟嘌呤的5 '侧链切割倾向于被抑制，但在G反应中不受抑制。
Adducts derived from the reaction of the benzo [a] pyrene metabolite model compound (+)-anfi-7| 8, 8a-dihydroxy-9a, 10a-epoxy-7, 8, 9, 10-tetrahydrobenzo [a] pyrene [(+)-BPDE] with the single-stranded oligodeoxynucleotide 5'-d (TATGCGTAT) were obtained according to direct synthesis techniques described earlier [Cosman, M., Ibanez, V., Geacintov, N. E., and Harvey, R. G.(1990) Carcinogenesis 11, 1667-1672]. Four major adducts, involving trans and cis addition (trans/cis adduct ratio* 4.5) of (+)-BPDE to the exocyclic amino groups of guanines G4 and Ge (the numbers denote the positions of the guanines counted from the 5'-side) were obtained. These adducts can be separated from one another by reverse-phase high-performance liquid chromatography methods. The site of BPDE binding on either 64 or G «can be determined from the electrophoresis band patterns on 20% polyacrylamide gels of the BPDE-modified oligonucleotides subjected to the G+ A and G Maxam-Gilbert strand cleavage reactions [Maxam, A. M., and Gilbert, W.(1980) Methods. Enzymol. 65, 499-560]. The electrophoresis gel band patterns are different for unmodifiedDNA and the two different BPDE-modified oligonucleotides because (1) the strand cleavage fragments bearing BPDE residues migrate slower than the corresponding fragments derived from the unmodified oligonucleotide and (2) strand cleavage tends to be inhibited on the 5'-sides of BPDE-modified guanines in the G+ A, but not the G reaction.