Antigenic analysis of Pseudomonas aeruginosa and Pseudomonas cepacia GroEL proteins and demonstration of a lipopolysaccharide-associated GroEL fraction in P. aeruginosa.
Quantitative crossed immunoelectrophoresis was used to evaluate the antigenic similarity of Pseudomonas aeruginosa and Pseudomonas cepacia GroEL proteins. We found that the two proteins showed 75% identity. By using a panel of monoclonal antibodies against the P. aeruginosa GroEL protein, we identified 10 monoclonal antibodies which cross-reacted with the P. cepacia GroEL protein and 21 monoclonal antibodies which recognized type-specific epitopes on the P. aeruginosa GroEL protein. In crossed immunoelectrophoresis two different fractions of GroEL reactive material could be resolved. These fractions showed a reaction of partial identity. Examination of the two immunoprecipitates by Western blotting, showed that both fractions consisted of anti-60 kDa GroEL reactive protein. One fraction, in addition, contained LPS with a characteristic 'ladder' reaction in modified Western blotting. We therefore conclude that this fraction represents a complex between LPS and GroEL.
Jensen
P
P
Department of Medical Microbiology, University of Copenhagen, Denmark.
Fomsgaard
A
A
Shand
G
G
Hindersson
P
P
Høiby
N
N
D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology; D000942:Antigens, Bacterial / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D001426:Bacterial Proteins / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D015153:Blotting, Western; D016956:Burkholderia cepacia / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D018834:Chaperonin 60; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000276:immunology; D006360:Heat-Shock Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007122:Immunoelectrophoresis; D008070:Lipopolysaccharides / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D018834:Chaperonin 60; D000939:Epitopes; D006360:Heat-Shock Proteins; D008070:Lipopolysaccharides