Quantitative determination of N-acetylglucosamine residues at the non-reducing ends of peptidoglycan chains by enzymic attachment of [14C]-D-galactose.
The ability of human milk galactosyltransferase to attach D-galactose residues quantitatively to the C-4 of N-acetylglucosamine moieties at the ends of oligosaccharides has been utilized for the specific labeling and quantitative determination of the chain length of the glycan moiety of the bacterial cell wall. The average polysaccharide chain length of the soluble, uncrosslinked peptidoglycan secreted by Micrococcus luteus cells on incubation with penicillin G was studied with this technique and found to be approximately 70 hexosamines long. Furthermore, the peptidoglycan chain length of Escherichia coli sacculi of different cell shapes and dimensions was determined both in rod-shaped cells and in filaments induced by temperature shift of a division mutant or by addition of cephalexin or nalidixic acid. The average chain length found in most of these sacculi was between 70 and 100 hexosamines long. Small spherical 'mini' cells had chain lengths similar to those of the isogenic rod-like cells.
D000117:Acetylglucosamine / Q000032:analysis*; D002250:Carbon Radioisotopes; D005260:Female; D005690:Galactose; D005700:Galactosyltransferases; D005944:Glucosamine / Q000031:analogs & derivatives*; D006595:Hexosamines / Q000032:analysis; D006801:Humans; D007553:Isotope Labeling / Q000379:methods; D008895:Milk, Human / Q000201:enzymology; D010457:Peptidoglycan; D011247:Pregnancy
D002250:Carbon Radioisotopes; D006595:Hexosamines; D010457:Peptidoglycan; D005700:Galactosyltransferases; D005944:Glucosamine; D000117:Acetylglucosamine; D005690:Galactose