Structure of the tetragonal surface virulence array protein and gene of Aeromonas salmonicida
| Autor: | S, Chu, S, Cavaignac, J, Feutrier, B M, Phipps, M, Kostrzynska, W W, Kay, T J, Trust |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
DNA
Bacterial Membrane Glycoproteins Base Sequence Virulence Factors Blotting Western Molecular Sequence Data Restriction Mapping Open Reading Frames Bacterial Proteins Genes Bacterial Escherichia coli Electrophoresis Polyacrylamide Gel Aeromonas Amino Acid Sequence Cloning Molecular Bacterial Outer Membrane Proteins Plasmids |
| Zdroj: | The Journal of biological chemistry. 266(23) |
| ISSN: | 0021-9258 |
| 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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