Structure of Bordetella pertussis peptidoglycan.

Abstract

Bordetella pertussis Tohama phases I and III were grown to the late-exponential phase in liquid medium containing [3H]diaminopimelic acid and treated by a hot (96 degrees C) sodium dodecyl sulfate extraction procedure. Washed sodium dodecyl sulfate-insoluble residue from phases I and III consisted of complexes containing protein (ca. 40%) and peptidoglycan (60%). Subsequent treatment with proteinase K yielded purified peptidoglycan which contained N-acetylglucosamine, N-acetylmuramic acid, alanine, glutamic acid, and diaminopimelic acid in molar ratios of 1:1:2:1:1 and less than 2% protein. Radiochemical analyses indicated that 3H added in diaminopimelic acid was present in peptidoglycan-protein complexes and purified peptidoglycan as diaminopimelic acid exclusively and that pertussis peptidoglycan was not O acetylated, consistent with it being degraded completely by hen egg white lysozyme. Muramidase-derived disaccharide peptide monomers and peptide-cross-linked dimers and higher oligomers were isolated by molecular-sieve chromatography; from the distribution of these peptidoglycan fragments, the extent of peptide cross-linking of both phase I and III peptidoglycan was calculated to be ca. 48%. Unambiguous determination of the structure of muramidase-derived peptidoglycan fragments by fast atom bombardment-mass spectrometry and tandem mass spectrometry indicated that the pertussis peptidoglycan monomer fraction was surprisingly homogeneous, consisting of greater than 95% N-acetylglucosaminyl-N-acetylmuramyl-alanyl-glutamyl-diaminopimelyl++ +-alanine.

• Authors: W J Folkening, W Nogami, S A Martin, R S Rosenthal
• Journal: Journal of bacteriology (J Bacteriol) Vol. 169 Issue 9 Pg. 4223-7 (Sep 1987) ISSN: 0021-9193 [Print] United States
• PMID: 2887547 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Amino Acids
• Peptidoglycan
• Diaminopimelic Acid
• Muramidase

Topics

• Acetylation
• Amino Acids (analysis)
• Bordetella pertussis (analysis, metabolism)
• Chromatography, High Pressure Liquid
• Chromatography, Paper
• Diaminopimelic Acid (analysis, metabolism)
• Mass Spectrometry
• Muramidase (metabolism)
• Peptidoglycan (analysis)