The structural basis for pyocin resistance in Neisseria gonorrhoeae lipooligosaccharides.
Pyocin resistance in a strain of Neisseria gonorrhoeae has been found to be associated with structural differences in the oligosaccharide moieties of the gonococcal outer membrane lipooligosaccharides (LOS). N. gonorrhoeae strain 1291 had been treated with several pyocins, usually lethal bacteriocins produced by Pseudomonas aeruginosa, and a series of surviving mutants were selected. The LOS of these pyocin-resistant mutants had altered electrophoretic mobilities in sodium dodecyl sulfate-polyacrylamide gels (Dudas, K. C., and Apicella, M. A. (1988) Infect. Immun. 56, 499-504). Structural analyses of the oligosaccharide portions of the wild-type (1291 wt) and five pyocin-resistant strains (1291a-e) by liquid secondary ion mass spectrometry, tandem mass spectrometry, and methylation analysis revealed that four of the mutant strains make oligosaccharides that differ from the wild-type LOS by successive saccharide deletions (1291a,c-e) and, in the oligosaccharide of 1291b, by the addition of a terminal Gal to the 1291c structure. The composition, sequence, and linkages of the terminal tetrasaccharide of the wild-type LOS are the same as the lacto-N-neotetraose terminus of the human paragloboside (Gal beta 1----4GlcNAc beta 1----3Gal beta 1----4Glc-ceramide), and both glycolipids bound the same monoclonal antibodies O6B4/3F11 that recognize this terminal epitope. None of the pyocin-resistant mutants bound this antibody. The 1291b LOS bound a monoclonal antibody that is specific for Gal alpha 1----4Gal beta 1----4Glc-ceramide (Pk glycosphingolipid) and shared a common composition, sequence, and linkages with this latter glycosphingolipid. Organisms that bound the anti-Pk monoclone occurred at the rate of approximately 1/750 among the wild-type parent strain. This structural information supports the conclusion that treatment with pyocin selects for mutants with truncated LOS structures and suggests that the oligosaccharides contained in the LOS of the wild-type strain and 1291b mimic those of human glycosphingolipids.
John
C M
CM
Department of Pharmaceutical Chemistry, University of California, San Francisco 94143.
Griffiss
J M
JM
Apicella
M A
MA
Mandrell
R E
RE
Gibson
B W
BW
D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D004352:Drug Resistance, Microbial; D004591:Electrophoresis, Polyacrylamide Gel; D005798:Genes, Bacterial; D007477:Ions; D008070:Lipopolysaccharides / Q000378:metabolism*; D013058:Mass Spectrometry; D008745:Methylation; D008969:Molecular Sequence Data; D009154:Mutation; D009344:Neisseria gonorrhoeae / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000378:metabolism; D011709:Pyocins / Q000494:pharmacology*; D011863:Radioimmunoassay
D007477:Ions; D008070:Lipopolysaccharides; D011709:Pyocins