Macromolecular organisation of recombinant Yersinia pestis F1 antigen and the effect of structure on immunogenicity.
Yersinia pestis, the causative organism of plague, produces a capsular protein (fraction 1 or F1 antigen) that is one of the major virulence factors of the bacterium. We report here the production, structural and immunological characterisation of a recombinant F1 antigen (rF1). The rF1 was purified by ammonium sulfate fractionation followed by FPLC Superose gel filtration chromatography. Using FPLC gel filtration chromatography and capillary electrophoresis, we have demonstrated that rF1 antigen exists as a multimer of high molecular mass. This multimer dissociates after heating in the presence of SDS and reassociation occurs upon the removal of SDS. Using circular dichroism, we have monitored the reassociation of monomeric rF1 into a multimeric form. Mice immunised with monomeric or multimeric rF1 develop similar immune responses, but mice immunised with monomeric rF1 were significantly less well protected against a challenge of 1 x 10(6) cfu of Y. pestis than mice immunised with multimeric rF1 (1/7 compared with 5/7). The significance of this result in terms of the structure and the function of rF1 is discussed.
Defence Evaluation and Research Agency, Salisbury, Wiltshire, UK.
J
J
Miller
E D
ED
Williamson
J H
JH
Lakey
M J
MJ
Pearce
S M
SM
Jones
R W
RW
Titball
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000737:chemistry* / Q000276:immunology* / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry* / Q000276:immunology* / Q000302:isolation & purification; D002942:Circular Dichroism; D005260:Female; D006358:Hot Temperature; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D010930:Plague / Q000517:prevention & control; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D012967:Sodium Dodecyl Sulfate; D015010:Yersinia pestis / Q000276:immunology*
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D011994:Recombinant Proteins; C086725:caf1 protein, Yersinia pestis; D012967:Sodium Dodecyl Sulfate