Complete amino acid sequence and location of Omp-28, an important immunogenic protein from Salmonella enterica serovar typhi.
Omp-28 isolated from Salmonella enterica serovar typhi presented a subunit molecular mass of 9,632 Da by MALDI-TOF MS. It was denatured, S-alkylated, and 1) directly submitted to Edman sequencing, 2) cleaved with CNBr, and 3) hydrolyzed either with endoproteinase Glu-C or Asp-N. The major CNBr peptide containing the C-terminal portion of Omp-28 was isolated by tricine-SDS-PAGE and electroblotted whereas Omp-28 enzymatic peptides were isolated by C18-RP-HPLC. All peptides were sequenced. This approach allowed the elucidation of the complete primary structure of Omp-28. Its amino acid sequence is identical to that deduced from part of the DNA of the "putative periplasmic transport protein" of either S. enterica serovar typhimurium and a multiple drug resistant S. enterica serovar typhi. Omp-28 homologous protein sequences were also deduced from Escherichia coli and Yersinia pestis genomic DNA. All proteins had their secondary structures predicted. Immunogold cytochemistry indicated that Omp-28 is found on the bacterium outer membrane.
Neves-Ferreira
Ana G C
AG
Departamento de Fisiologia e Farmacodinâmica, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
de Andrade
Carlos M
CM
Vannier-Santos
Marcos A
MA
Perales
Jonas
J
Nascimento
Hilton J
HJ
da Silva Junior
José G
JG
D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D016680:Genome, Bacterial; D006090:Gram-Negative Bacteria / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D012485:Salmonella typhi / Q000235:genetics* / Q000276:immunology / Q000648:ultrastructure; D020539:Sequence Analysis, Protein; D012689:Sequence Homology, Nucleic Acid
D001425:Bacterial Outer Membrane Proteins; C116185:OMP-28 protein, Salmonella typhi