Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2014 Dec 16:4:7494.
doi: 10.1038/srep07494.

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

Daniel Kühner  1 , Mark Stahl  2 , Dogan D Demircioglu  3 , Ute Bertsche  1
Affiliations

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

Daniel Kühner et al. Sci Rep. .

Abstract

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.

PubMed Disclaimer

Figures

Figure 1
Figure 1. Muropeptide profile of S. aureus SA113 obtained by UPLC and UPLC/MS.
(a) Muropeptide profile of S. aureus SA113 obtained by reversed phase UPLC. (b) TIC of UPLC/MS analysis of S. aureus SA113 obtained by reversed-phase UPLC coupled to MS. Masses of indicated peaks are shown in table 1 and table S1 including molecule composition and proposed sum formula.
Figure 2
Figure 2. Muropeptide profile of E. coli Nissle 1917 obtained by UPLC and UPLC/MS.
(a)Muropeptide profile of E. coli Nissle 1917 obtained by reversed phase UPLC. (b) TIC of UPLC/MS analysis of E. coli Nissle 1917 obtained by reversed-phase UPLC coupled to MS. Masses of indicated peaks are shown in table 2 and table S2 including molecule composition and proposed sum formula.
Figure 3
Figure 3. Schematic structure of muropeptides of S. aureus and E. coli.
The basic structure consists of the disaccharide (GlcNAc-MurNAc) with an adjacent stem peptide (L-Ala – D-iGlx – L-Lys – D-Ala – D-Ala) with Glx being either Gln or Glu. Our model organism S. aureus, harbors also a five-glycine interpeptide bridge branching from the L-Lys, which constitutes indirect cross-links between two adjacent stem peptides, as it is depicted in the multimeric muropeptide. The C at position 6 of the MurNAc can be O-acetylated.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Schleifer K. H. & Kandler O. Peptidoglycan types of bacterial cell walls and their taxonomic implications. Bacteriol Rev 36, 407–477 (1972). - PMC - PubMed
    1. Salton M. R. J. & Horne R. W. Studies of the bacterial cell wall. 2. Methods of preparation and some properties of cell walls. Biochimica et Biophysica Acta 7, 177–197 (1951). - PubMed
    1. Glauner B. Separation and quantification of muropeptides with high-performance liquid chromatography. Anal Biochem 172, 451–464 (1988). - PubMed
    1. de Jonge B. L., Chang Y. S., Gage D. & Tomasz A. Peptidoglycan composition of a highly methicillin-resistant Staphylococcus aureus strain. The role of penicillin binding protein 2A. J Biol Chem 267, 11248–11254 (1992). - PubMed
    1. Desmarais S. M., Cava F., de Pedro M. A. & Huang K. C. Isolation and Preparation of Bacterial Cell Walls for Compositional Analysis by Ultra Performance Liquid Chromatography. JoVE e51183, 10.3791/51183 (2014). - DOI - PMC - PubMed

Publication types

MeSH terms