Localizing Peptidoglycan Synthesis in Helicobacter pylori using Clickable Metabolic Probes.

Localizing Peptidoglycan Synthesis in Helicobacter pylori using Clickable Metabolic Probes.
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DOI:
10.1002/cpz1.80
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发表时间:
2021-04
期刊:
Current protocols
影响因子:
--
通讯作者:
Salama NR
Salama NR
中科院分区:
其他
文献类型:
--
作者:
Taylor JA;Santiago CC;Gray J;Wodzanowski KA;DeMeester KE;Biboy J;Vollmer W;Grimes CL;Salama NR

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细菌细胞壁由肽聚糖(PG)组成,为细胞提供结构完整性,并负责大多数细菌的细胞形状。在这里,我们提出了使用可点击的PG特异性糖MurNAc-alk作为代谢探针来研究细胞壁的工具。在这里,我们提出了一个新的反应途径,产生MurNAc-alk。我们还包括标记幽门螺杆菌PG合成的协议,使用LC-MS/MS确定标记的muropeptide的身份,样品制备的细胞标记的倍增时间的一小部分,和可视化使用3D结构照明显微镜。
The bacterial cell wall, composed of peptidoglycan (PG), provides structural integrity for the cell and is responsible for cell shape in most bacteria. Here we present tools to study the cell wall using a clickable PG-specific sugar, MurNAc-alk, as a metabolic probe. Here we present a new reaction pathway for generating MurNAc-alk. We also include protocols for labeling PG synthesis in Helicobacter pylori, determining the identity of the labeled muropeptides using LC-MS/MS, sample preparation of cells labeled for a short fraction of the doubling time, and visualization using 3D structured illumination microscopy.