Structural characterization of the homotropic cooperative binding of azamulin to human cytochrome P450 3A5.

IF 0.1 4区 文学 0 LITERATURE, SLAVIC
SLAVIC AND EAST EUROPEAN JOURNAL Pub Date : 2022-05-01 Epub Date: 2022-04-06 DOI:10.1016/j.jbc.2022.101909
Mei-Hui Hsu, Eric F Johnson
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引用次数: 0

Abstract

Cytochrome P450 3A4 and 3A5 catalyze the metabolic clearance of a large portion of therapeutic drugs. Azamulin is used as a selective inhibitor for 3A4 and 3A5 to define their roles in metabolism of new chemical entities during drug development. In contrast to 3A4, 3A5 exhibits homotropic cooperativity for the sequential binding of two azamulin molecules at concentrations used for inhibition. To define the underlying sites and mechanisms for cooperativity, an X-ray crystal structure of 3A5 was determined with two azamulin molecules in the active site that are stacked in an antiparallel orientation. One azamulin resides proximal to the heme in a pose similar to the 3A4-azamulin complex. Comparison to the 3A5 apo structure indicates that the distal azamulin in 3A5 ternary complex causes a significant induced fit that excludes water from the hydrophobic surfaces of binding cavity and the distal azamulin, which is augmented by the stacking interaction with the proximal azamulin. Homotropic cooperativity was not observed for the binding of related pleuromutilin antibiotics, tiamulin, retapamulin, and lefamulin, to 3A5, which are larger and unlikely to bind in the distal site in a stacked orientation. Formation of the 3A5 complex with two azamulin molecules may prevent time-dependent inhibition that is seen for 3A4 by restricting alternate product formation and/or access of reactive intermediates to vulnerable protein sites. These results also contribute to a better understanding of sites for cooperative binding and the differential structural plasticity of 3A5 and 3A4 that contribute to differential substrate and inhibitor binding.

氮杂环丁烷与人类细胞色素 P450 3A5 同向协同结合的结构特征。
细胞色素 P450 3A4 和 3A5 催化了大部分治疗药物的代谢清除。阿扎木林被用作 3A4 和 3A5 的选择性抑制剂,以确定它们在药物开发过程中对新化学实体代谢的作用。与 3A4 不同的是,3A5 在抑制浓度下与两个氮杂环戊烷分子依次结合时表现出同向合作性。为了确定合作性的基本位点和机制,我们测定了 3A5 的 X 射线晶体结构,其活性位点中有两个以反平行方向堆叠的氮杂氨分子。其中一个氮杂氨分子位于血红素近端,其姿势与 3A4 氮杂氨分子复合物相似。与 3A5 apo 结构的比较表明,3A5 三元复合物中的远端偶氮嘧啶会产生明显的诱导配合,将水排除在结合腔和远端偶氮嘧啶的疏水表面之外,而与近端偶氮嘧啶的堆叠相互作用又增强了这种诱导配合。在相关的胸腺嘧啶抗生素(tiamulin、retapamulin 和 lefamulin)与 3A5 的结合中没有观察到同向合作作用,这些抗生素较大,不太可能以堆叠方向结合到远端位点。3A5 与两个氮氨嘧啶分子形成复合物,可能会限制交替产物的形成和/或反应性中间产物进入脆弱的蛋白质位点,从而防止 3A4 出现的时间依赖性抑制。这些结果还有助于更好地理解合作结合位点以及 3A5 和 3A4 不同的结构可塑性,它们有助于不同的底物和抑制剂结合。
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来源期刊
SLAVIC AND EAST EUROPEAN JOURNAL
SLAVIC AND EAST EUROPEAN JOURNAL LITERATURE, SLAVIC-
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