title: Mycobacterial lipid II is composed of a complex mixture of modified muramyl and peptide moieties linked to decaprenyl phosphate.
abstract: Structural analysis of compounds identified as lipid I and II from Mycobacterium smegmatis demonstrated that the lipid moiety is decaprenyl phosphate; thus, M. smegmatis is the first bacterium reported to utilize a prenyl phosphate other than undecaprenyl phosphate as the lipid carrier involved in peptidoglycan synthesis. In addition, mass spectrometry showed that the muropeptides from lipid I are predominantly N-acetylmuramyl-L-alanine-D-glutamate-meso-diaminopimelic acid-D-alanyl-D-alanine, whereas those isolated from lipid II form an unexpectedly complex mixture in which the muramyl residue and the pentapeptide are modified singly and in combination. The muramyl residue is present as N-acetylmuramic acid, N-glycolylmuramic acid, and muramic acid. The carboxylic functions of the peptide side-chains of lipid II showed three types of modification, with the dominant one being amidation. The preferred site for amidation is the free carboxyl group of the meso-diaminopimelic acid residue. Diamidated species were also observed. The carboxylic function of the terminal D-alanine of some molecules is methylated, as are all three carboxylic acid functions of other molecules. This study represents the first structural analysis of mycobacterial lipid I and II and the first report of extensive modifications of these molecules. The observation that lipid I was unmodified strongly suggests that the lipid II intermediates of M. smegmatis are substrates for a variety of enzymes that introduce modifications to the sugar and amino acid residues prior to the synthesis of peptidoglycan.
authors.affiliation: Mycobacterial Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523, USA.
authors.forename: Sebabrata
authors.initials: S
authors.lastname: Mahapatra
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Yagi
authors.forename: John T
authors.initials: JT
authors.lastname: Belisle
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Espinosa
authors.forename: Preston J
authors.initials: PJ
authors.lastname: Hill
authors.forename: Michael R
authors.initials: MR
authors.lastname: McNeil
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Brennan
authors.forename: Dean C
authors.initials: DC
authors.lastname: Crick
mesh_terms: D002473:Cell Wall / Q000737:chemistry*; D009161:Mycobacterium / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010457:Peptidoglycan / Q000737:chemistry / Q000378:metabolism; D011106:Polyisoprenyl Phosphates / Q000378:metabolism*; D014538:Uridine Diphosphate N-Acetylmuramic Acid / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D010457:Peptidoglycan; D011106:Polyisoprenyl Phosphates; D014538:Uridine Diphosphate N-Acetylmuramic Acid; C069249:muramyl-NAc-(pentapeptide)pyrophosphoryl-undecaprenol; C068697:decaprenyl phosphate