Identification of epitope and surface-exposed domains of Shigella flexneri invasion plasmid antigen D (IpaD).
Transport and surface expression of the invasion plasmid antigens (Ipa proteins) is an essential trait in the pathogenicity of Shigella spp. In addition to the type III protein secretion system encoded by the mxi/spa loci on the large virulence plasmid, transport of IpaB and IpaC into the surrounding medium is modulated by IpaD. To characterize the structural topography of IpaD, the Geysen epitope-mapping system was used to identify epitopes recognized by surface-reactive monoclonal and polyclonal antibodies produced against purified recombinant IpaD or synthetic IpaD peptides. Surface-exposed epitopes of IpaD were confined to the first 180 amino acid residues, whereas epitopes in the carboxyl-terminal half were not exposed on the Shigella surface. By using convalescent-phase sera from 10 Shigella flexneri-infected monkeys, numerous epitopes were mapped within a surface-exposed region of IpaD between amino acid residues 14 and 77. Epitopes were also identified in the carboxyl-terminal half of IpaD with a few convalescent-phase sera. Comparison of IpaD epitope sequences with Salmonella SipD sequences indicated that very similar epitopes may exist in the carboxyl-terminal region of each protein whereas the IpaD epitopes in the surface-exposed amino-terminal region were unique for the Shigella protein. Although the IpaD and SipD homologs may play similar roles in transport, the dominant serum antibody response to IpaD is against the unique region of this protein exposed on the surface of the pathogen.
Turbyfill
K R
KR
Department of Enteric Infections, Walter Reed Army Institute of Research, Washington, DC 20307, USA.
Mertz
J A
JA
Mallett
C P
CP
Oaks
E V
EV
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000942:Antigens, Bacterial / Q000276:immunology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000276:immunology*; D018604:Epitope Mapping; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids; D017433:Protein Structure, Secondary; D011817:Rabbits; D012763:Shigella flexneri / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C095770:IpaD protein, Shigella flexneri