Biogenesis of Yersinia pestis PsaA in Recombinant Attenuated Salmonella Typhimurium Vaccine (RASV) Strain
| Autor: | Torres-Escobar, Ascención, Juárez-Rodríguez, María Dolores, Curtiss, Roy |
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| Jazyk: | angličtina |
| Rok vydání: | 2009 |
| Předmět: |
Salmonella typhimurium
Antigens Bacterial Plague Vaccine Vaccines Synthetic Binding Sites Base Sequence Genetic Vectors Molecular Sequence Data Serine Endopeptidases Membrane Proteins Protein Sorting Signals Article Bacterial Proteins Mutagenesis Site-Directed Aspartic Acid Endopeptidases Humans Amino Acid Sequence Sequence Alignment Sequence Deletion |
| Popis: | Yersinia pestis PsaA is an adhesin important for the establishment of bacterial infection. PsaA synthesis requires the products of the psaEFABC genes. Here, by prediction analysis, we identified a PsaA signal sequence with two signal peptidase (SPase) cleavage sites, type-I and type-II (SPase-I and SPase-II). By Edman degradation and site-directed mutagenesis, the precise site for one of these Spase-I PsaA cleavage sites was located between alanine and serine at positions 31 and 32, respectively. Yersinia pestis psaA expression and the role of the PsaB and PsaC proteins were evaluated in recombinant attenuated Salmonella Typhimurium vaccine strains. PsaA was detected in total extracts as a major 15-kDa (mature) and 18-kDa (unprocessed) protein bands. PsaA synthesis was not altered by a DeltaA31-DeltaS32 double-deletion mutation. In contrast, the synthesis of PsaA (DeltaA31-DeltaS32) in Y. pestis and delivery to the supernatant was decreased. Otherwise, substitution of the amino acid cysteine at position 26 by valine involved in the SPase-II cleavage site did not show any effect on the secretion of PsaA in Salmonella and Yersinia. These results help clarify the secretion pathway of PsaA for the possible development of vaccines against Y. pestis. |
| Databáze: | OpenAIRE |
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