Published March 24, 2015 | Version v1
Journal article Open

From cells to muropeptide structures in 24 h: Peptidoglycan mapping by UPLC-MS

Creators

  • 1. ScientificProtocols.org

Description

Peptidoglycan (PGN) is ubiquitous in nearly all bacterial species. The PGN sacculus protects the cells against their own internal turgor making PGN one of the most important targets for antibacterial treatment. Within the last sixty years PGN composition has been intensively studied by various methods. The breakthrough was the application of HPLC technology on the analysis of muropeptides. However, preparation of pure PGN relied on a very time consuming method of about one week. We established a purification protocol for both Gram-positive and Gram-negative bacteria which can be completely performed in plastic reaction tubes yielding pure muropeptides within 24 hours. The muropeptides can be analyzed by UPLC-MS, allowing their immediate determination. This new rapid method provides the feasibility to screen PGN composition even in high throughput, making it a highly useful tool for basic research as well as for the pharmaceutical industry.

Files

protocol.md

Files (16.4 kB)

Name Size Download all
md5:baa45efe296f9bd4eb9f4f53f55d8798
16.4 kB Preview Download