Investigating the Roles of Active Site Residues in Mycobacterium tuberculosis Indole-3-glycerol Phosphate Synthase, a Potential Target for Antitubercular Agents

IF 3.8 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY ACS Bio & Med Chem Au Pub Date : 2023-07-26 DOI:10.1021/acsbiomedchemau.3c00029
David W. Konas, Sarah Cho, Oshane D. Thomas, Maryum M. Bhatti, Katherine Leon Hernandez, Cinthya Moran, Hedda Booter, Thomas Candela, Joseph Lacap, Paige McFadden, Savannah van den Berg, Alyssa M. Welter, Ashley Peralta, Cheryl A. Janson, Jaclyn Catalano and Nina M. Goodey*, 
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Abstract

Mycobacterium tuberculosis drug resistance is emerging and new drug targets are needed. Tryptophan biosynthesis is necessary for M. tuberculosis replication and virulence. Indole-3-glycerol phosphate synthase (IGPS) catalyzes a step in M. tuberculosis tryptophan biosynthesis and has been suggested as a potential anti-infective target, but our understanding of this enzyme is limited. To aid in inhibitor design and gain a greater mechanistic picture of this enzyme, there is a need to understand the roles of active site amino acids in ligand binding and catalysis. In this work, we explored the roles of conserved active site amino acids Glu57, Lys59, Lys119, Glu168, and Glu219. Mutation of each to Ala results in loss of all detectable activity. The Glu57Gln, Lys59Arg, Lys119Arg, Glu168Gln, and Glu219Asp mutations result in large activity losses, while Glu219Gln has enhanced activity. Analysis of the enzymatic data yields the following main conclusions: (A) Lys119 is the likely catalytic acid in the CdRP ring closure step. (B) Glu168 stabilizes a charged reaction intermediate and may also be the catalytic base. (C) Glu57, Glu219, and Lys119 form a closely arranged triad in which Glu57 and Glu219 modulate the pKa of Lys119, and thus overall activity. This increased understanding of inter- and intramolecular interactions and demonstration of the highly coordinated nature of the M. tuberculosis IGPS active site provide new mechanistic information and guidance for future work with this potential new drug target.

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结核分枝杆菌吲哚-3-甘油磷酸合酶活性位点残基在抗结核药物潜在靶点中的作用研究
结核分枝杆菌耐药性正在出现,需要新的药物靶点。色氨酸的生物合成是结核分枝杆菌复制和毒力所必需的。吲哚-3-甘油磷酸合成酶(IGPS)催化结核分枝杆菌色氨酸生物合成的一个步骤,并被认为是一个潜在的抗感染靶点,但我们对这种酶的了解有限。为了帮助抑制剂的设计并获得这种酶的更深入的机制,需要了解活性位点氨基酸在配体结合和催化中的作用。在这项工作中,我们探索了保守的活性位点氨基酸Glu57、Lys59、Lys119、Glu168和Glu219的作用。每个到Ala的突变导致所有可检测活性的丧失。Glu57Gln、Lys59Arg、Lys119Arg、Glu168Gln和Glu219Asp突变导致大的活性损失,而Glu219Gln具有增强的活性。酶数据的分析得出以下主要结论:(A)Lys119可能是CdRP环闭合步骤中的催化酸。(B) Glu168稳定带电的反应中间体,也可以是催化碱。(C) Glu57、Glu219和Lys119形成紧密排列的三联体,其中Glu57和Glu219调节Lys119的pKa,从而调节整体活性。这增加了对分子间和分子内相互作用的理解,并证明了结核分枝杆菌IGPS活性位点的高度协调性,为未来研究这一潜在的新药靶点提供了新的机制信息和指导。
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ACS Bio & Med Chem Au
ACS Bio & Med Chem Au 药物、生物、化学-
CiteScore
4.10
自引率
0.00%
发文量
0
期刊介绍: ACS Bio & Med Chem Au is a broad scope open access journal which publishes short letters comprehensive articles reviews and perspectives in all aspects of biological and medicinal chemistry. Studies providing fundamental insights or describing novel syntheses as well as clinical or other applications-based work are welcomed.This broad scope includes experimental and theoretical studies on the chemical physical mechanistic and/or structural basis of biological or cell function in all domains of life. It encompasses the fields of chemical biology synthetic biology disease biology cell biology agriculture and food natural products research nucleic acid biology neuroscience structural biology and biophysics.The journal publishes studies that pertain to a broad range of medicinal chemistry including compound design and optimization biological evaluation molecular mechanistic understanding of drug delivery and drug delivery systems imaging agents and pharmacology and translational science of both small and large bioactive molecules. Novel computational cheminformatics and structural studies for the identification (or structure-activity relationship analysis) of bioactive molecules ligands and their targets are also welcome. The journal will consider computational studies applying established computational methods but only in combination with novel and original experimental data (e.g. in cases where new compounds have been designed and tested).Also included in the scope of the journal are articles relating to infectious diseases research on pathogens host-pathogen interactions therapeutics diagnostics vaccines drug-delivery systems and other biomedical technology development pertaining to infectious diseases.
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