Purification, characterization and sequence analysis of Omp50,a new porin isolated from Campylobacter jejuni.
A novel pore-forming protein identified in Campylobacter was purified by ion-exchange chromatography and named Omp50 according to both its molecular mass and its outer membrane localization. We observed a pore-forming ability of Omp50 after re-incorporation into artificial membranes. The protein induced cation-selective channels with major conductance values of 50-60 pS in 1 M NaCl. N-terminal sequencing allowed us to identify the predicted coding sequence Cj1170c from the Campylobacter jejuni genome database as the corresponding gene in the NCTC 11168 genome sequence. The gene, designated omp50, consists of a 1425 bp open reading frame encoding a deduced 453-amino acid protein with a calculated pI of 5.81 and a molecular mass of 51169.2 Da. The protein possessed a 20-amino acid leader sequence. No significant similarity was found between Omp50 and porin protein sequences already determined. Moreover, the protein showed only weak sequence identity with the major outer-membrane protein (MOMP) of Campylobacter, correlating with the absence of antigenic cross-reactivity between these two proteins. Omp50 is expressed in C. jejuni and Campylobacter lari but not in Campylobacter coli. The gene, however, was detected in all three species by PCR. According to its conformation and functional properties, the protein would belong to the family of outer-membrane monomeric porins.
Bolla
J M
JM
CJF 96-06 INSERM, Faculté de Médecine la Timone, Université de la Méditerranée, 27 Bd Jean Moulin, 13385 Marseille Cedex 05, France. Jean-Michel.Bolla@medecine.univ-mrs.fr
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Pagès
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D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D015153:Blotting, Western; D016123:Campylobacter jejuni / Q000737:chemistry* / Q000235:genetics*; D002463:Cell Membrane Permeability; D002852:Chromatography, Ion Exchange; D003429:Cross Reactions / Q000276:immunology; D003902:Detergents; D004553:Electric Conductivity; D004591:Electrophoresis, Polyacrylamide Gel; D005798:Genes, Bacterial / Q000235:genetics; D007106:Immune Sera; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D018272:Porins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D011487:Protein Conformation; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D001426:Bacterial Proteins; D003902:Detergents; D007106:Immune Sera; D008051:Lipid Bilayers; D018272:Porins; C095080:major outer membrane protein, Campylobacter jejuni