Crystal Structure of the N-Acetylmuramic Acid α-1-Phosphate (MurNAc-α1-P) Uridylyltransferase MurU, a Minimal Sugar Nucleotidyltransferase and Potential Drug Target Enzyme in Gram-negative Pathogens
Crystal Structure of the N-Acetylmuramic Acid α-1-Phosphate (MurNAc-α1-P) Uridylyltransferase MurU, a Minimal Sugar Nucleotidyltransferase and Potential Drug Target Enzyme in Gram-negative Pathogens
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DOI：
10.1074/jbc.m114.620989
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发表时间：
2015-04-24
影响因子：
4.8
通讯作者：
Stehle, Thilo
中科院分区：
文献类型：
--
作者：
Renner-Schneck, Michaela;Hinderberger, Isabel;Stehle, Thilo
N-乙酰胞壁酸 α-1-磷酸 (MurNAc-α 1-P) 尿苷酰转移酶 MurU 催化尿苷二磷酸 (UDP)-MurNAc 的合成，尿苷二磷酸 (UDP)-MurNAc 是细菌肽聚糖细胞壁的重要前体。 MurU 是最近在革兰氏阴性细菌中发现的细胞壁回收途径的一部分，该途径绕过 UDP-MurNAc 的一般从头生物合成，并有助于对针对 UDP-MurNAc 从头生物合成的抗生素磷霉素产生高度内在耐药性。为了深入了解底物结合和特异性，我们以高分辨率解析了恶臭假单胞菌 MurU 在天然状态和配体结合状态下的晶体结构。在这些结构的帮助下，确定了关键的酶-底物相互作用，使 MurNAc-α 1-P 能够与 MurU 的活性位点紧密结合。 MurU 结构定义了一般核苷酸转移酶活性所需的“最小结构域”。它们还为 MurU 抑制剂的化学设计提供了结构基础，该抑制剂可以作为针对多重耐药革兰氏阴性病原体的联合治疗中的新药。
The N-acetylmuramic acid alpha-1-phosphate (MurNAc-alpha 1-P) uridylyltransferase MurU catalyzes the synthesis of uridine diphosphate (UDP)-MurNAc, a crucial precursor of the bacterial peptidoglycan cell wall. MurU is part of a recently identified cell wall recycling pathway in Gram-negative bacteria that bypasses the general de novo biosynthesis of UDP-MurNAc and contributes to high intrinsic resistance to the antibiotic fosfomycin, which targets UDP-MurNAc de novo biosynthesis. To provide insights into substrate binding and specificity, we solved crystal structures of MurU of Pseudomonas putida in native and ligand-bound states at high resolution. With the help of these structures, critical enzyme-substrate interactions were identified that enable tight binding of MurNAc-alpha 1-P to the active site of MurU. The MurU structures define a "minimal domain" required for general nucleotidyltransferase activity. They furthermore provide a structural basis for the chemical design of inhibitors of MurU that could serve as novel drugs in combination therapy against multidrug-resistant Gram-negative pathogens.