Purification, location, and immunological characterization of the iron-regulated high-molecular-weight proteins of the highly pathogenic yersiniae.
We have previously shown that under iron limitation, different Yersinia species synthesize new polypeptides. Two of them, the high-molecular-weight proteins (HMWPs), are expressed only by the highly pathogenic strains. In the present study, the HMWPs from Y. enterocolitica serovar O:8 were purified by gel filtration, and specific antibodies were obtained. Using these antibodies, we show that the two polypeptides were synthesized de novo during iron starvation and that they were found essentially in the bacterial outer membrane fractions, although the majority of the molecules were not exposed on the cell surface. We also demonstrate that the two proteins had common epitopes and that the HMWPs of the high-virulence-phenotype species Y. pestis, Y. pseudotuberculosis, and Y. enterocolitica serovar O:8 (a strain different from the one used to purify the proteins) are antigenically related. The less pathogenic and nonpathogenic strains did not exhibit cross-reacting material, suggesting that these strains do not synthesize even an altered form of the HMWPs.
Carniel
E
E
Unité d'Ecologie Bacterienne, Institut Pasteur, Paris, France.
Antoine
J C
JC
Guiyoule
A
A
Guiso
N
N
Mollaret
H H
HH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D002451:Cell Compartmentation; D002850:Chromatography, Gel; D003429:Cross Reactions; D003470:Culture Media; D004220:Disulfides; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007501:Iron / Q000494:pharmacology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D010957:Plasmids; D015007:Yersinia / Q000032:analysis / Q000235:genetics / Q000472:pathogenicity*
D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D003470:Culture Media; D004220:Disulfides; D007501:Iron