Establishment of the experimental procedure for prediction of conjugation capacity in mutant UGT1A1
Establishment of the experimental procedure for prediction of conjugation capacity in mutant UGT1A1
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DOI：
10.1371/journal.pone.0225244
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发表时间：
2019-11-15
期刊：
影响因子：
3.7
通讯作者：
Sakaeda, Toshiyuki
中科院分区：
文献类型：
--
作者：
Takaoka, Yutaka;Takeuchi, Atsuko;Sakaeda, Toshiyuki
UDP-葡萄糖醛酸基转移酶1A 1（UGT 1A 1）是一种在内质网膜中发现的酶，据报道可具有大量导致葡萄糖醛酸化能力降低的氨基酸取代。例如，当患者接受CPT-11（伊立替康）时，如在癌症化疗中，药物不良反应是由UGT 1A 1中的氨基酸取代引起的。我们先前发现，当胆红素的羟基残基朝向UDP-葡萄糖醛酸时，对接的程度与体外缀合能力相关。在这项研究中，我们分析了突变体UGT 1A 1的构象通过结构优化与水和脂质双层，而不是在真空中，我们在我们以前的研究中使用的优化。然后，我们推导出一个数学模型，该模型可以通过使用计算机模拟底物对接的结果和UGT 1A 1对对乙酰氨基酚和17 β-雌二醇葡萄糖醛酸化的体外分析结果来预测突变UGT 1A 1的缀合能力。该实验程序表明，其他突变体UGT 1A 1与胆红素或SN-38的计算机模拟结合能力与报告的体外结合能力相似。我们的结果表明，本文所述的实验程序可以正确预测突变体UGT 1A 1和任何底物的缀合能力。
UDP-glucuronosyltransferase 1A1 (UGT1A1) is an enzyme that is found in the endoplasmic reticulum membrane and can reportedly have a large number of amino acid substitutions that result in the reduction of glucuronidation capacity. For example, adverse drug reactions when patients receive CPT-11 (irinotecan) such as in cancer chemotherapy are caused by amino acid substitutions in UGT1A1. We previously found that the extent of the docking when the hydroxyl residue of bilirubin was oriented toward UDP-glucuronic acid correlated with in vitro conjugation capacity. In this study, we analyzed the conformation of mutant UGT1A1s by means of structural optimization with water and lipid bilayers instead of the optimization in vacuo that we used in our previous study. We then derived a mathematical model that can predict the conjugation capacities of mutant UGT1A1s by using results of substrate docking in silico and results of in vitro analysis of glucuronidation of acetaminophen and 17 beta-estradiol by UGT1A1s. This experimental procedure showed that the in silico conjugation capacities of other mutant UGT1A1s with bilirubin or SN-38 were similar to reported in vitro conjugation capacities. Our results suggest that this experimental procedure described herein can correctly predict the conjugation capacities of mutant UGT1A1s and any substrate.