The TraT lipoprotein as a vehicle for the transport of foreign antigenic determinants to the cell surface of Escherichia coli K12: structure-function relationships in the TraT protein.
The TraT protein is a surface-exposed lipoprotein, specified by plasmids of the IncF group, that mediates serum resistance and surface exclusion. The structure and function of the TraT protein determined by plasmid R6-5 was probed by genetic insertion of a foreign antigenic determinant, the C3 epitope of polio virus, at residues 61, 125, 180, 200 or 216 of the protein. The chimaeric proteins were transported to the outer membrane and, in three cases, immunoassays with an anti-C3 monoclonal antibody indicated that the C3 epitope was exposed on the cell surface. Three of the hybrids, with insertions at residues 125, 180 and 200, assembled into the trypsin-resistant oligomeric form characteristic of the wild-type protein, which suggested that these regions are not involved in TraT subunit:subunit interactions. Additionally, the hybrid protein carrying the C3 epitope at position 180 functioned in a genetic suppression assay and retained partial surface-exclusion activity. Thus, its localization, folding and organization does not appear to be grossly altered from that of the wild-type protein. Applications of the protein for the transport of foreign antigenic determinants to the cell surface are discussed.
Taylor
I M
IM
Department of Biochemistry, University of Southampton, UK.
Harrison
J L
JL
Timmis
K N
KN
O'Connor
C D
CD
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D001483:Base Sequence; D001692:Biological Transport; D002213:Capsid / Q000276:immunology*; D002462:Cell Membrane / Q000276:immunology; D002678:Chimera; D004251:DNA Transposable Elements; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000378:metabolism*; D004926:Escherichia coli / Q000276:immunology / Q000378:metabolism*; D029968:Escherichia coli Proteins; D006168:Guinea Pigs; D008969:Molecular Sequence Data; D010957:Plasmids; D017955:Poliovirus / Q000276:immunology*; D015183:Restriction Mapping; D013329:Structure-Activity Relationship
D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D004251:DNA Transposable Elements; D000939:Epitopes; D029968:Escherichia coli Proteins; C036297:TraT protein, E coli