title: Structural characterization of peptidoglycan muropeptides by matrix-assisted laser desorption ionization mass spectrometry and postsource decay analysis.
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.
authors.affiliation: Department of Biochemistry, Michigan State University, East Lansing 48824, USA.
authors.forename: N
authors.initials: N
authors.lastname: Xu
authors.forename: Z H
authors.initials: ZH
authors.lastname: Huang
authors.forename: B L
authors.initials: BL
authors.lastname: de Jonge
authors.forename: D A
authors.initials: DA
authors.lastname: Gage
mesh_terms: D000595:Amino Acid Sequence; D008248:Lysostaphin; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009112:Muramic Acids / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D010457:Peptidoglycan / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D009112:Muramic Acids; D010455:Peptides; D010457:Peptidoglycan; D008248:Lysostaphin