Structural Characterization of Neisseria gonorrhoeae Bacterial Peroxidase-Insights into the Catalytic Cycle of Bacterial Peroxidases.
Structural Characterization of Neisseria gonorrhoeae Bacterial Peroxidase-Insights into the Catalytic Cycle of Bacterial Peroxidases.
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DOI：
10.3390/ijms24076246
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发表时间：
2023-03-26
影响因子：
5.6
通讯作者：
Pauleta, Sofia R.
中科院分区：
文献类型：
--
作者：
Nobrega, Claudia S.;Carvalho, Ana Luisa;Romao, Maria Joao;Pauleta, Sofia R.
关键词：
淋病奈瑟菌是一种人类特有的致病细菌，可导致淋病，这是一种性传播疾病。细菌过氧化物酶是一种存在于这种细菌周质中的酶，它能使细胞免受过氧化氢的毒害，是这种有机体抵御外源和内源性氧化应激的主要防御措施之一。异源产生的38 kDa细菌过氧化物酶在pH 6.0下以混合价、活化态结晶，晶体用叠氮浸泡，得到该酶家族的第一个叠氮抑制结构。该酶结合外源配体，如氰化物和叠氮化物，这也抑制了催化活性，通过协调活性部位的P血红素铁，并与其底物过氧化氢竞争。测得氰化物和叠氮化物的抑制常数分别为0.4±0.1µM和41±5 mm。咪唑还通过与P和E血红素结合而以更复杂的机制结合和抑制该酶，这改变了最新的血红素的还原潜力。根据目前报道的结构，重新讨论了细菌过氧化物酶的催化循环。抑制研究和抑制酶的晶体结构构成了寻找和开发以该酶为靶点的抑制剂的第一个平台，作为可能的治疗淋病奈瑟菌的新策略。
Neisseria gonorrhoeae is an obligate human pathogenic bacterium responsible for gonorrhea, a sexually transmitted disease. The bacterial peroxidase, an enzyme present in the periplasm of this bacterium, detoxifies the cells against hydrogen peroxide and constitutes one of the primary defenses against exogenous and endogenous oxidative stress in this organism. The 38 kDa heterologously produced bacterial peroxidase was crystallized in the mixed-valence state, the active state, at pH 6.0, and the crystals were soaked with azide, producing the first azide-inhibited structure of this family of enzymes. The enzyme binds exogenous ligands such as cyanide and azide, which also inhibit the catalytic activity by coordinating the P heme iron, the active site, and competing with its substrate, hydrogen peroxide. The inhibition constants were estimated to be 0.4 ± 0.1 µM and 41 ± 5 mM for cyanide and azide, respectively. Imidazole also binds and inhibits the enzyme in a more complex mechanism by binding to P and E hemes, which changes the reduction potential of the latest heme. Based on the structures now reported, the catalytic cycle of bacterial peroxidases is revisited. The inhibition studies and the crystal structure of the inhibited enzyme comprise the first platform to search and develop inhibitors that target this enzyme as a possible new strategy against N. gonorrhoeae.
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影响因子：
2.9
作者：
Echalier, Aude;Brittain, Thomas;Watmough, Nicholas J.
通讯作者：
Watmough, Nicholas J.
影响因子：
3.9
作者：
Dumortier, C;Meyer, TE;Cusanovich, MA
通讯作者：
Cusanovich, MA
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
影响因子：
2.9
作者：
Chen, YR;Deterding, LJ;Mason, RP
通讯作者：
Mason, RP
影响因子：
8
作者：
Hadarovich, Anna;Chakravarty, Devlina;Vakser, Ilya A.
通讯作者：
Vakser, Ilya A.