Inhibition of biological activities of the aerobactin receptor protein in rough strains of Escherichia coli by polyclonal antiserum raised against native protein.
The aerobactin iron-uptake system of plasmid ColV-K30, genetically isolated from other plasmid determinants by molecular cloning, was sufficient to restore full virulence in a mouse peritonitis model to a clinical Escherichia coli isolate, D551 (O78:H-), whose resident aerobactin-encoding ColV plasmid had been lost by curing. Antiserum was raised in rabbits against live E. coli K12 cells expressing the outer-membrane aerobactin receptor protein and absorbed with an isogenic strain lacking the receptor. This antiserum inhibited binding of aerobactin, cloacin DF13 and bacteriophage B74K to the native protein in whole E. coli K12 bacteria expressing the receptor, or in membranes prepared from such organisms. However, it did not react with the native receptor protein in several wild strains unless lipopolysaccharide was first removed by treatment with trichloroacetic acid, nor did it protect mice in experimental infections with strain D551. Antisera raised in rabbits against partially or fully denatured forms of the aerobactin receptor reacted only in assays involving denatured protein; they showed no inhibition of the biological activities of the native receptor.
Roberts
M
M
Department of Genetics, University of Leicester, UK.
Wooldridge
K G
KG
Gavine
H
H
Kuswandi
S I
SI
Williams
P H
PH
D000818:Animals; D001425:Bacterial Outer Membrane Proteins; D004926:Escherichia coli / Q000472:pathogenicity*; D004927:Escherichia coli Infections / Q000235:genetics / Q000276:immunology* / Q000517:prevention & control; D007106:Immune Sera / Q000276:immunology; D015151:Immunoblotting; D051379:Mice; D008807:Mice, Inbred BALB C; D011971:Receptors, Immunologic / Q000276:immunology / Q000378:metabolism*; D014774:Virulence / Q000235:genetics / Q000276:immunology
D001425:Bacterial Outer Membrane Proteins; D007106:Immune Sera; D011971:Receptors, Immunologic; C034506:aerobactin receptor