Surface hydrophobics mediate functional dimerization of CYP121A1 of Mycobacterium tuberculosis.
Surface hydrophobics mediate functional dimerization of CYP121A1 of Mycobacterium tuberculosis.
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DOI：
10.1038/s41598-020-79545-y
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发表时间：
2021-01-11
影响因子：
4.6
通讯作者：
Estrada DF
中科院分区：
文献类型：
--
作者：
Kumar A;Campomizzi CS;Jay N;Ferguson S;Scheffler EJ;Lioi J;Tu C;Qu J;Simons C;Estrada DF
结核病是由致病细菌结核分枝杆菌(Mtb)引起的，仍然是世界范围内感染致死的主要原因。结核分枝杆菌基因组编码不成比例的20种细胞色素P450酶，其中基本酶细胞色素P450 121A1(CYP121A1)仍然是药物设计努力的目标。CYP121A1介导了酪氨酸二肽环-L-酪氨酸-L-酪氨酸(CYY)的苯酚偶联反应。在这项工作中，二聚的结构和功能被作为一个被忽视的特征的CYP121A1功能进行了研究。这项研究表明，CYP121A1二聚体通过分子间接触在远端表面形成，并由暴露在溶剂中的疏水残基网络介导。通过定点突变破坏CYP121A1二聚体会导致cYY的部分特异性丧失，导致催化作用下降约75%。19F标记和酶FG-loop的核磁共振还与蛋白质对接相结合，开发了具有功能的CYP121A1二聚体的工作模型。这些结果表明，同源二聚体界面参与底物选择性，代表了细胞色素P450底物结合的一种新范例，同时也为开发新型抗结核药物中的相关药物靶点提供了重要的机制见解。
Tuberculosis is caused by the pathogenic bacterium Mycobacterium tuberculosis (Mtb) and remains the leading cause of death by infection world-wide. The Mtb genome encodes a disproportionate number of twenty cytochrome P450 enzymes, of which the essential enzyme cytochrome P450 121A1 (CYP121A1) remains a target of drug design efforts. CYP121A1 mediates a phenol coupling reaction of the tyrosine dipeptide cyclo-L-Tyr-L-Tyr (cYY). In this work, a structure and function investigation of dimerization was performed as an overlooked feature of CYP121A1 function. This investigation showed that CYP121A1 dimers form via intermolecular contacts on the distal surface and are mediated by a network of solvent-exposed hydrophobic residues. Disruption of CYP121A1 dimers by site-directed mutagenesis leads to a partial loss of specificity for cYY, resulting in an approximate 75% decrease in catalysis. 19F labeling and nuclear magnetic resonance of the enzyme FG-loop was also combined with protein docking to develop a working model of a functional CYP121A1 dimer. The results obtained suggest that participation of a homodimer interface in substrate selectivity represents a novel paradigm of substrate binding in CYPs, while also providing important mechanistic insight regarding a relevant drug target in the development of novel anti-tuberculosis agents.
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影响因子：
14.8
作者：
Kozakov D;Hall DR;Xia B;Porter KA;Padhorny D;Yueh C;Beglov D;Vajda S
通讯作者：
Vajda S
影响因子：
14.8
作者：
通讯作者：
--
DOI：
10.1073/pnas.1616198114
发表时间：
2017-01-17
影响因子：
11.1
作者：
Sevrioukova, Irina F.;Poulos, Thomas L.
通讯作者：
Poulos, Thomas L.
影响因子：
3
作者：
Byrne, Sean T.;Denkin, Steven M.;Zhang, Ying
通讯作者：
Zhang, Ying
影响因子：
3.7
作者：
Rose-Sperling, Dania;Mai Anh Tran;Hellmich, Ute A.
通讯作者：
Hellmich, Ute A.