Structure of the tetragonal surface virulence array protein and gene of Aeromonas salmonicida
| Autor: | Trevor J. Trust, William W. Kay, J Feutrier, S Cavaignac, M Kostrzynska, B M Phipps, Shijian Chu |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
biology
Structural gene Protein primary structure Cell Biology medicine.disease_cause biology.organism_classification Trypsin Biochemistry Molecular biology Aeromonas salmonicida Open reading frame medicine Molecular Biology Escherichia coli Peptide sequence Protein secondary structure medicine.drug |
| Zdroj: | Journal of Biological Chemistry. 266:15258-15265 |
| ISSN: | 0021-9258 |
| DOI: | 10.1016/s0021-9258(18)98611-4 |
| Popis: | The paracrystalline surface protein array of the pathogenic bacterium Aeromonas salmonicida is a primary virulence factor with novel binding capabilities. The species-specific structural gene (vapA) for this array protein (A-protein) was cloned into lambda gt11 but was unstable when expressed in Escherichia coli, undergoing an 816-base pair deletion due to a 21-base pair direct repeat within the gene. However, the gene was stable in cosmid pLA2917 as long as expression was poor. A-protein was located in the cytoplasmic, inner membrane and periplasmic fractions in E. coli. The DNA sequence revealed a 1,506-base pair open reading frame encoding a protein consisting of a 21-amino acid signal peptide, and a 481-residue 50,778 molecular weight protein containing considerable secondary structure. When assembled into a paracrystalline protein array on Aeromonas the cell surface A-protein was totally refractile to cleavage by trypsin, but became trypsin sensitive when disassembled. Trypsin cleavage of the isolated protein provided evidence that both the NH2- and COOH-terminal regions form distinct structural domains, consistent with three-dimensional ultrastructural evidence. The NH2-terminal 274-residue domain remained refractile to trypsin activity. This segment connects by a trypsin and CNBr-sensitive 78-residue linker region to a COOH-terminal 129-residue fragment which could apparently refold into a partially trypsin-resistant structure after cleavage at residue 323. |
| Databáze: | OpenAIRE |
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