Vaught J B, Lee M S, Shayman M A, Thissen M R, King C M
Chem Biol Interact. 1981 Feb;34(1):109-24. doi: 10.1016/0009-2797(81)90095-8.
Reaction of N-hydroxy-2-aminofluorene (N-OH-AF) with rRNA at pH 5.0 decreased the molecular weight of the polynucleotide. Toluene-soluble aryl derivatives were released on hydrolysis of fluorenylamine- and biphenylamine-substituted RNA by treatment with venom phosphodiesterase and alkaline phosphatase. These data suggested that arylhydroxylamines, activated by incubation at pH 5.0 or by enzymatic O-acetylation, might react with the phosphate group of RNA to give unstable phosphate triesters. Spontaneous hydrolysis of these triesters would result in cleavage of the polynucleotide chain. Further enzymatic hydrolysis of the phosphate esters would yield nonpolar arylamine derivatives. Enzymatically degraded 4-aminobiphenyl(ABP)-RNA adducts were examined by high performance liquid chromatography (HPLC) for the presence of a putative phosphorylated adduct. Synthetic standards of the C-8-guanosine monophosphate-ABP adduct (ABP-GMP) and o-aminobiphenyl-O-phosphate were used as markers in the analysis of the digested RNA. A phosphate adduct of ABP was undetectable by these methods. The data also indicated that the ABP-GMP formed in the acyltransferase-mediated binding of N-hydroxy-4-acetylaminobiphenyl (N-OH-AABP) to RNA is readily degraded during the enzymatic digestion of the RNA adduct.
在pH 5.0条件下，N-羟基-2-氨基芴（N-OH-AF）与rRNA的反应降低了多核苷酸的分子量。用蛇毒磷酸二酯酶和碱性磷酸酶处理芴胺和联苯胺取代的RNA后，水解产物释放出甲苯可溶的芳基衍生物。这些数据表明，在pH 5.0孵育或酶促O-乙酰化激活的芳基羟胺可能与RNA的磷酸基团反应生成不稳定的磷酸三酯。这些三酯的自发水解会导致多核苷酸链的断裂。磷酸酯的进一步酶促水解会产生非极性芳胺衍生物。通过高效液相色谱（HPLC）检测酶促降解的4-氨基联苯（ABP）-RNA加合物中是否存在假定的磷酸化加合物。在分析消化后的RNA时，使用C-8-鸟苷单磷酸-ABP加合物（ABP-GMP）和邻氨基联苯-O-磷酸的合成标准品作为标志物。通过这些方法无法检测到ABP的磷酸加合物。数据还表明，在酰基转移酶介导的N-羟基-4-乙酰氨基联苯（N-OH-AABP）与RNA结合过程中形成的ABP-GMP在RNA加合物的酶促消化过程中很容易降解。
