title: Dynamics of the exposure of epitopes on OmpF, an outer membrane protein of Escherichia coli.
abstract: The OmpF protein is the major outer membrane trimeric porin of Escherichia coli B. The exposure of several cell-surface-exposed epitopes, that are recognized by various monoclonal antibodies directed against the protein, is investigated. Kinetic analyses show that two epitopes (E18 and E19) appear early during the in-vivo assembly on the folded monomer, just after the removal of the signal peptide, and are conserved in the native trimer. The trimerization that immediately follows or occurs in conjunction with the folding of monomers exposes another antigenic site (E21) at the surface of metastable forms. The binding of nascent lipopolysaccharide promotes the conversion of the heat-modifiable intermediate to a stable trimer and ensures the exposure of E20, E1, E3, E4 and E7. Late epitopes, E1, E3, E4 and E7 are only detected in the outer membrane fraction. These results suggest that different steps induce the sequential exposure of native antigenic sites. The detection of these epitopes depends on conformational changes occurring during the OmpF insertion into the outer membrane.
authors.affiliation: Centre de Biochimie et de Biologie Moléculaire, Centre National de la Recherche Scientifique, Marseille, France.
authors.forename: D
authors.initials: D
authors.lastname: Fourel
authors.forename: S
authors.initials: S
authors.lastname: Mizushima
authors.forename: J M
authors.initials: JM
authors.lastname: Pagès
mesh_terms: D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D004926:Escherichia coli / Q000378:metabolism*; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D011487:Protein Conformation; D021382:Protein Sorting Signals / Q000032:analysis / Q000378:metabolism; D013462:Sulfur Radioisotopes
chemical_list: D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D000939:Epitopes; D021382:Protein Sorting Signals; D013462:Sulfur Radioisotopes; D008715:Methionine