Analysis of the peptidoglycan structure of Bacillus subtilis endospores.
Peptidoglycan was prepared from purified Bacillus subtilis spores of wild-type and several mutant strains. Digestion with muramidase resulted in cleavage of the glycosidic bonds adjacent to muramic acid replaced by peptide or alanine side chains but not the bonds adjacent to muramic lactam. Reduction of the resulting muropeptides allowed their separation by reversed-phase high-pressure liquid chromatography. The structures of 20 muropeptides were determined by amino acid and amino sugar analysis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. In wild-type spores, 50% of the muramic acid had been converted to the lactam and 75% of these lactam residues were spaced regularly at every second muramic acid position in the glycan chains. Single L-alanine side chains were found on 25% of the muramic acid residues. The remaining 25% of the muramic acid had tetrapeptide or tripeptide side chains, and 11% of the diaminopimelic acid in these side chains was involved in peptide cross-links. Analysis of spore peptidoglycan produced by a number of mutants lacking proteins involved in cell wall metabolism revealed structural changes. The most significant changes were in the spores of a dacB mutant which lacks the sporulation-specific penicillin-binding protein 5*. In these spores, only 46% of the muramic acid was in the lactam form, 12% had L-alanine side chains, and 42% had peptide side chains containing diaminopimelic acid, 29% of which was involved in cross-links.
Popham
D L
DL
Department of Biochemistry, University of Connecticut Health Center, Farmington 06030-3305, USA.
Helin
J
J
Costello
C E
CE
Setlow
P
P
D000117:Acetylglucosamine / Q000032:analysis; D000596:Amino Acids / Q000032:analysis; D001412:Bacillus subtilis / Q000737:chemistry* / Q000235:genetics; D001426:Bacterial Proteins; D002240:Carbohydrate Sequence; D002352:Carrier Proteins / Q000235:genetics; D002851:Chromatography, High Pressure Liquid; D003960:Diaminopimelic Acid / Q000032:analysis; D006020:Glycopeptides / Q000737:chemistry; D006602:Hexosyltransferases; D008969:Molecular Sequence Data; D009112:Muramic Acids / Q000032:analysis; D002267:Muramoylpentapeptide Carboxypeptidase / Q000235:genetics; D009154:Mutation; D009842:Oligopeptides / Q000737:chemistry; D046915:Penicillin-Binding Proteins; D010457:Peptidoglycan / Q000737:chemistry*; D010458:Peptidyl Transferases; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013171:Spores, Bacterial / Q000737:chemistry*
D000596:Amino Acids; D001426:Bacterial Proteins; D002352:Carrier Proteins; D006020:Glycopeptides; D009112:Muramic Acids; D009842:Oligopeptides; D046915:Penicillin-Binding Proteins; D010457:Peptidoglycan; C031651:N-acetylmuramic acid; D003960:Diaminopimelic Acid; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D002267:Muramoylpentapeptide Carboxypeptidase; D000117:Acetylglucosamine