Complete Amino Acid Sequence and Location of Omp-28, an Important Immunogenic Protein from Salmonella enterica serovar typhi |
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Authors: | Ana G C Neves-Ferreira Carlos M de Andrade Marcos A Vannier-Santos Jonas Perales Hilton J Nascimento José G da Silva Junior |
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Institution: | Departamento de Fisiologia e Farmacodinamica, Instituto Oswaldo Cruz, Funda??o Oswaldo Cruz, Rio de Janeiro, Brazil. |
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Abstract: | 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. |
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Keywords: | Amino acid sequence Omp-28 outer membrane protein Salmonella enterica serovar typhi |
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