Catalytic process of anhydro-N-acetylmuramic acid kinase from Pseudomonas aeruginosa.
Catalytic process of anhydro-N-acetylmuramic acid kinase from Pseudomonas aeruginosa.
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DOI：
10.1016/j.jbc.2023.105198
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发表时间：
2023-10
影响因子：
4.8
通讯作者：
Mobashery, Shahriar
中科院分区：
文献类型：
--
作者：
El-Araby, Amr M.;Jimenez-Faraco, Eva;Feltzer, Rhona;Martin-Garcia, Jose M.;Karri, Bhaskara Rao;Ramachandran, Balajee;Kim, Choon;Fisher, Jed F.;Hermoso, Juan A.;Mobashery, Shahriar
细菌细胞膜是细菌与其环境接触并免受其环境影响的结构。该包膜内有一种保守的肽聚糖聚合物，它赋予细胞包膜形状和强度。构建、重塑和回收这种交联聚合物的化学成分的酶促过程是抗生素的卓越目标和新兴抗生素结构的探索目标。我们报告了一种此类酶——铜绿假单胞菌脱水-N-乙酰胞壁酸（anhNAM）激酶（AnmK）的全面动力学和结构分析。 AnmK 是该病原体肽聚糖回收途径中的一种酶。它催化 anhNAM 的水解开环与伴随的 ATP 依赖性磷酰基转移的配对。 AnmK 的 anhNAM 和 ATP 底物遵循随机顺序动力学机制。对四种不同结构（apo AnmK、AnmK:AMPPNP、AnmK:AMPPNP:anhNAM 和 AnmK:ATP:anhNAM）的晶体分析表明，两种底物以底物亚位点的非门控构象独立进入活性位点，其中蛋白环充当 anhNAM 结合的门。催化作用发生在酶的闭合构象状态下。我们使用 ATP 模拟分子在晶体学上观察到这种状态。二聚 AnmK 的显着 X 射线结构揭示了催化前和催化后三元配合物。计算模拟结合高分辨率 X 射线结构揭示了完整的催化循环。我们进一步报告说，与 WT 菌株相比，anmK 基因被破坏的铜绿假单胞菌菌株对 β-内酰胺亚胺培南更敏感。这些观察结果使 AnmK 能够理解肽聚糖回收、抗生素敏感性和细菌毒力之间的关系。
The bacterial cell envelope is the structure with which the bacterium engages with, and is protected from, its environment. Within this envelop is a conserved peptidoglycan polymer which confers shape and strength to the cell envelop. The enzymatic processes that build, remodel, and recycle the chemical components of this cross-linked polymer are preeminent targets of antibiotics and exploratory targets for emerging antibiotic structures. We report a comprehensive kinetic and structural analysis for one such enzyme, the Pseudomonas aeruginosa anhydro-N-acetylmuramic acid (anhNAM) kinase (AnmK). AnmK is an enzyme in the peptidoglycan-recycling pathway of this pathogen. It catalyzes the pairing of hydrolytic ring opening of anhNAM with concomitant ATP-dependent phosphoryl transfer. AnmK follows a random-sequential kinetic mechanism with respect to its anhNAM and ATP substrates. Crystallographic analyses of four distinct structures (apo AnmK, AnmK:AMPPNP, AnmK:AMPPNP:anhNAM, and AnmK:ATP:anhNAM) demonstrate that both substrates enter the active site independently in an ungated conformation of the substrate subsites, with protein loops acting as gates for anhNAM binding. Catalysis occurs within a closed conformational state for the enzyme. We observe this state crystallographically using ATP-mimetic molecules. A remarkable X-ray structure for dimeric AnmK sheds light on the precatalytic and postcatalytic ternary complexes. Computational simulations in conjunction with the high-resolution X-ray structures reveal the full catalytic cycle. We further report that a P. aeruginosa strain with disrupted anmK gene is more susceptible to the β-lactam imipenem compared to the WT strain. These observations position AnmK for understanding the nexus among peptidoglycan recycling, susceptibility to antibiotics, and bacterial virulence.
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影响因子：
4.6
作者：
Byun B;Mahasenan KV;Dik DA;Marous DR;Speri E;Kumarasiri M;Fisher JF;Hermoso JA;Mobashery S
通讯作者：
Mobashery S
AnmK 样激酶的计算分析：真菌细胞壁代谢的新见解
影响因子：
2
作者：
Dai, Jianghong;Qu, Hong;Zhang, Hongxun
通讯作者：
Zhang, Hongxun
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：
4.8
作者：
Bacik, John-Paul;Whitworth, Garrett E.;Mark, Brian L.
通讯作者：
Mark, Brian L.
影响因子：
5.7
作者：
Dominguez-Gil, Teresa;Lee, Mijoon;Hermoso, Juan A.
通讯作者：
Hermoso, Juan A.