Department of Microbiology, Institute of Biomedical Science, University of São Paulo, Av. Prof Lineu Prestes, 1374, CEP 05508-000 São Paulo, SP, Brazil.
Department of Microbiology, Institute of Biomedical Science, University of São Paulo, Av. Prof Lineu Prestes, 1374, CEP 05508-000 São Paulo, SP, Brazil; Department of Chemistry. The University of Warwick, Coventry CV4 7AL, UK.
J Struct Biol. 2023 Jun;215(2):107957. doi: 10.1016/j.jsb.2023.107957. Epub 2023 Mar 20.
The cytoplasmatic biosynthesis of the stem peptide from the peptidoglycan in bacteria involves six steps, which have the role of three ATP-dependent Mur ligases that incorporate three consecutive amino acids to a substrate precursor. MurE is the last Mur ligase to incorporate a free amino acid. Although the structure of MurE from Mycobacterium tuberculosis (MtbMurE) was determined at 3.0 Å, the binding mode of meso-Diaminopimelate (m-DAP) and the effect of substrate absence is unknown. Herein, we show the structure of MurE from M. thermoresistibile (MthMurE) in complex with ADP and m-DAP at 1.4 Å resolution. The analysis of the structure indicates key conformational changes that the substrate UDP-MurNAc-L-Ala-D-Glu (UAG) and the free amino acid m-DAP cause on the MthMurE conformation. We observed several movements of domains or loop regions that displace their position in order to perform enzymatic catalysis. Since MthMurE has a high similarity to MtbMurE, this enzyme could also guide strategies for structure-based antimicrobial discovery to fight against tuberculosis or other mycobacterial infections.
细菌中肽聚糖衍生的茎肽的细胞质生物合成涉及六个步骤,其中三个 ATP 依赖性 Mur 连接酶的作用是将三个连续的氨基酸连接到一个底物前体上。MurE 是最后一个将游离氨基酸掺入到产物中的 Mur 连接酶。尽管结核分枝杆菌(MtbMurE)的 MurE 结构已在 3.0Å 下确定,但 m-DAP 的结合模式以及缺乏底物的影响尚不清楚。在这里,我们展示了耐热分枝杆菌(MthMurE)与 ADP 和 m-DAP 复合物的 MurE 结构,分辨率为 1.4Å。结构分析表明,底物 UDP-MurNAc-L-Ala-D-Glu(UAG)和游离氨基酸 m-DAP 会引起 MthMurE 构象的关键构象变化。我们观察到几个结构域或环区域的运动,它们改变了位置以进行酶催化。由于 MthMurE 与 MtbMurE 具有高度相似性,因此该酶也可以为基于结构的抗菌药物发现提供指导,以对抗结核病或其他分枝杆菌感染。