Pseudomonas aeruginosa lipopolysaccharide: evidence that the O side chains and common antigens are on the same molecule.
We investigated whether Pseudomonas aeruginosa produces two distinct lipopolysaccharides (LPS) containing either serologically variable O side chains or a neutral polysaccharide common antigen, designated A bands, that reacts with monoclonal antibody (MAb) E87. Immunoprecipitation of LPS and free O side chains with O-side-chain-specific antibodies or MAb E87 resulted in coprecipitation of both polysaccharides when antibody of either specificity was employed. Chromatography of LPS and free O side chains in a disaggregating deoxycholate buffer indicated the two polysaccharide antigens cochromatograph when eluates were analyzed by sensitive and specific enzyme-linked immunosorbent assay inhibitions. The LPS from a mutant of strain PAO1 that lacks polymerized O side chains but retains the common antigen eluted in fractions containing smaller LPS molecules, indicating the necessity of polymerized O side chains for elution in early fractions containing large LPS monomers. A phosphomannomutase mutant of P. aeruginosa PAO1 makes a rough LPS lacking both O side chains and common antigen but still produces a small (< 6-kDa) common antigen component detectable in cell lysates. Introduction of the cloned pmm gene into this strain restored production of a smooth LPS expressing large MAb E87-reactive common antigen. Destruction with NaOH of O side chains on recombinant LPS molecules eluting early from the molecular sieve column resulted in a shift of the MAb E87-reactive antigen to the late-eluting fractions. These results indicate that on most P. aeruginosa LPS molecules, O side chains and neutral polysaccharide common antigens are both present.
Hatano
K
K
Channing Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115.
Goldberg
J B
JB
Pier
G B
GB
D000907:Antibodies, Bacterial / Q000276:immunology; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D007535:Isomerases / Q000235:genetics; D008070:Lipopolysaccharides / Q000276:immunology* / Q000302:isolation & purification; D019081:O Antigens; D017875:Phosphotransferases (Phosphomutases); D011135:Polysaccharides, Bacterial / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011233:Precipitin Tests; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000276:immunology*; D013045:Species Specificity; D013211:Staphylococcus aureus / Q000276:immunology
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000939:Epitopes; D008070:Lipopolysaccharides; D019081:O Antigens; D011135:Polysaccharides, Bacterial; D007535:Isomerases; D017875:Phosphotransferases (Phosphomutases); C052799:phosphomannomutase