Fries R W, Bohlken D P, Murch B P, Leidal K G, Plapp B V
Arch Biochem Biophys. 1983 Aug;225(1):110-5. doi: 10.1016/0003-9861(83)90012-7.
A series of esters of adenosine 5'-monophosphate with ethyl, propyl, or hexyl moieties substituted at the omega-position with chlorine or bromine were prepared. The compounds were competitive inhibitors of horse liver alcohol dehydrogenase with respect to coenzyme, NAD+, and had inhibition (dissociation) constants in the range of 40 to 260 microM at pH 8.0, 25 degrees C. The bromoalkyl esters were designed to be active-site-directed inactivators and were chemically reactive as tested with the model compound 4-(p-nitrobenzyl)pyridine. Yeast alcohol dehydrogenase was inactivated by the bromohexyl analog by an active-site-directed mechanism, with a Ki = 1.5 mM and a pseudo-bimolecular rate constant of 0.03 M-1 S-1, which is 150 times larger than the bimolecular rate constant for inactivation by 2-bromoethanol. However, the rates of inactivation of other dehydrogenases treated with 10 mM concentrations of these compounds were generally slower than with the simpler reagent, 2-bromoethanol. Thus, the reactive functional group attached to the AMP moiety may not be properly oriented for affinity labeling of these dehydrogenases. The bromoalkyl esters may be useful for inactivating other enzymes.
制备了一系列5'-单磷酸腺苷的酯类化合物，其乙酯、丙酯或己酯的ω位被氯或溴取代。这些化合物在辅酶NAD⁺方面是马肝醇脱氢酶的竞争性抑制剂，在pH 8.0、25℃时抑制（解离）常数范围为40至260微摩尔。溴代烷基酯被设计为活性位点导向的失活剂，并且如用模型化合物4-(对硝基苄基)吡啶测试的那样具有化学反应性。酵母醇脱氢酶被溴代己基类似物通过活性位点导向机制失活，Ki = 1.5毫摩尔，假双分子速率常数为0.03 M⁻¹ s⁻¹，这比2-溴乙醇失活的双分子速率常数大150倍。然而，用10毫摩尔浓度的这些化合物处理其他脱氢酶的失活速率通常比用更简单的试剂2-溴乙醇慢。因此，连接到AMP部分的反应性功能基团可能没有正确定向用于这些脱氢酶的亲和标记。溴代烷基酯可能可用于使其他酶失活。
