Structural characterization of peptidoglycan muropeptides by matrix-assisted laser desorption ionization mass spectrometry and postsource decay analysis

Analytical Biochemistry
N XuD A Gage

Abstract

In this study we report the development of matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS)-based methods for the structural characterization of muropeptides derived from peptidoglycan. Prior to analysis, peptidoglycan samples were subjected to enzymatic digestion with muramidase and the resulting muropeptides were purified by HPLC. A new matrix, 5-chloro-2-mercaptobenzothiazole, was employed for the MALDI-MS analysis. The results have demonstrated that sub-picomole to femtomole detection can be achieved in both positive mode and negative mode, allowing unambiguous determination of the molecular masses of monomeric and oligomeric muropeptides. Structural information from monomeric muropeptides was obtained by further postsource decay (PSD) analysis. Fragmentation patterns in positive mode and negative mode PSD were complementary for the elucidation of the peptide chain sequence. Lysostaphin digestion was also incorporated with MALDI mass mapping analysis for determination of peptide chain cross-linking patterns of muropeptide oligomers from Staphylococcus aureus strains.

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Citations

Jul 17, 2004·Microbial Drug Resistance : MDR : Mechanisms, Epidemiology, and Disease·Anatoly SeverinAlexander Tomasz
Sep 18, 2002·Antimicrobial Agents and Chemotherapy·Boudewijn L M De JongeNaxing Xu
Oct 17, 2006·Journal of Bacteriology·Catherine EckertStéphane Mesnage
Nov 21, 2008·Rapid Communications in Mass Spectrometry : RCM·Junjie Gao, Carolyn J Cassady
Jun 26, 1999·The Journal of Biological Chemistry·K SieradzkiA Tomasz
Jun 12, 2003·The Journal of Biological Chemistry·Aude AntignacIvo G Boneca
Sep 25, 2003·The Journal of Biological Chemistry·Zheng-Ming WangRoman Dziarski
Apr 13, 2021·Current Protocols·Jennifer A TaylorNina R Salama
Jul 20, 2019·Microbiology Spectrum·Rita Sobral, Alexander Tomasz

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Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.

Bacterial Cell Wall Structure

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.