结核分枝杆菌吡嗪酰胺酶的晶体结构：对天然和获得性吡嗪酰胺耐药性的深入了解。
Crystal structure of the pyrazinamidase of Mycobacterium tuberculosis: insights into natural and acquired resistance to pyrazinamide.
机构信息
Faculté de Médecine Pitié-Salpêtrière, UPMC Université Paris 6, ER 5, EA1541, Bactériologie-Hygiène, Paris, France.
出版信息
PLoS One. 2011 Jan 24;6(1):e15785. doi: 10.1371/journal.pone.0015785.
Abstract
Pyrazinamidase (PncA) activates the first-line antituberculous drug pyrazinamide into pyrazinoic acid. The crystal structure of the Mycobacterium tuberculosis PncA protein has been determined, showing significant differences in the substrate binding cavity when compared to the pyrazinamidases from Pyrococcus horikoshii and Acinetobacter baumanii. In M. tuberculosis, this region was found to hold a Fe(2+) ion coordinated by one aspartate and three histidines, one of them corresponding to His57 which is replaced by Asp in Mycobacterium bovis, a species naturally resistant to pyrazinamide. The binding cavity also contains a Cys138-Asp8-Lys96 motif evocating a cysteine-based catalytic mechanism. Mutants have been constructed and investigated by kinetic and thermal shift assays, highlighting the importance of protein folding and thermal stability in the pyrazinamidase activity.
摘要
吡嗪酰胺酶（PncA）能够将一线抗结核药物吡嗪酰胺转化为吡嗪酸。结核分枝杆菌 PncA 蛋白的晶体结构已经被确定，与来自 Pyrococcus horikoshii 和 Acinetobacter baumanii 的吡嗪酰胺酶相比，其底物结合腔存在显著差异。在结核分枝杆菌中，该区域被发现含有一个由一个天冬氨酸和三个组氨酸配位的 Fe（2+）离子，其中一个对应于 His57，而 His57 在天然对吡嗪酰胺具有抗性的牛分枝杆菌中被天冬氨酸取代。结合腔还包含一个 Cys138-Asp8-Lys96 基序，提示了基于半胱氨酸的催化机制。通过动力学和热位移测定法构建和研究了突变体，突出了蛋白质折叠和热稳定性在吡嗪酰胺酶活性中的重要性。
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本文引用的文献
1
All-atom empirical potential for molecular modeling and dynamics studies of proteins.
J Phys Chem B. 1998 Apr 30;102(18):3586-616. doi: 10.1021/jp973084f.
2
Kinetics and inhibition of nicotinamidase from Mycobacterium tuberculosis.
Biochemistry. 2010 Nov 9;49(44):9613-9. doi: 10.1021/bi1011157.
3
Insight to pyrazinamide resistance in Mycobacterium tuberculosis by molecular docking.
Bioinformation. 2009 Aug 18;4(1):24-9. doi: 10.6026/97320630004024.
4
Peruvian and globally reported amino acid substitutions on the Mycobacterium tuberculosis pyrazinamidase suggest a conserved pattern of mutations associated to pyrazinamide resistance.
Infect Genet Evol. 2010 Mar;10(2):346-9. doi: 10.1016/j.meegid.2009.11.016. Epub 2009 Dec 4.
5
Mutations found in the pncA gene of Mycobacterium tuberculosis in clinical pyrazinamide-resistant isolates from a local region of China.
J Int Med Res. 2009 Sep-Oct;37(5):1430-5. doi: 10.1177/147323000903700517.
6
Characterisation of pncA mutations in clinical Mycobacterium tuberculosis isolates in New Zealand.
Pathology. 2009;41(6):582-4. doi: 10.1080/00313020903071587.
7
Specificity and mechanism of Acinetobacter baumanii nicotinamidase: implications for activation of the front-line tuberculosis drug pyrazinamide.
Angew Chem Int Ed Engl. 2009;48(48):9176-9. doi: 10.1002/anie.200903407.
8
Mycobacterium tuberculosis genotypic diversity in pyrazinamide-resistant isolates of Iran.
Microb Drug Resist. 2009 Dec;15(4):251-6. doi: 10.1089/mdr.2009.0066.
9
Assessing compound binding to the Eg5 motor domain using a thermal shift assay.
Anal Biochem. 2009 Sep 1;392(1):59-69. doi: 10.1016/j.ab.2009.05.044. Epub 2009 Jun 2.
10
Phaser crystallographic software.
J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674. doi: 10.1107/S0021889807021206. Epub 2007 Jul 13.
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