title: From cells to muropeptide structures in 24 h: peptidoglycan mapping by UPLC-MS.
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.
authors.affiliation: Eberhard Karls University Tuebingen, IMIT - Microbial Genetics, Waldhaeuser Str. 70/8, 72076 Tuebingen, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Kühner
authors.affiliation: Eberhard Karls University Tuebingen, Center for Plant Molecular Biology, Central Facilities, Auf der Morgenstelle 32, 72076 Tuebingen, Germany.
authors.forename: Mark
authors.initials: M
authors.lastname: Stahl
authors.affiliation: Eberhard Karls University Tuebingen, IMIT - Microbial Genetics, Auf der Morgenstelle 28, 72076 Tuebingen, Germany.
authors.forename: Dogan D
authors.initials: DD
authors.lastname: Demircioglu
authors.affiliation: Eberhard Karls University Tuebingen, IMIT - Microbial Genetics, Waldhaeuser Str. 70/8, 72076 Tuebingen, Germany.
authors.forename: Ute
authors.initials: U
authors.lastname: Bertsche
mesh_terms: D002473:Cell Wall; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004926:Escherichia coli / Q000737:chemistry*; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry*; D010457:Peptidoglycan / Q000032:analysis / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013211:Staphylococcus aureus / Q000737:chemistry*
chemical_list: D010446:Peptide Fragments; D010457:Peptidoglycan