Isolation and identification of a putative porcine transferrin receptor from Actinobacillus pleuropneumoniae biotype 1.
Each of two affinity isolation methods, the first based on biotinylated porcine transferrin plus streptavidin-agarose, and the second on Sepharose-coupled porcine transferrin, followed by SDS-PAGE, allowed the isolation and identification of two potential porcine-transferrin-binding polypeptides (approximately 64 kDa and 99 kDa) from total membranes of Actinobacillus pleuropneumoniae grown under iron-restricted conditions. Both polypeptides were iron-repressible and were identified as potential receptor candidates as they were not isolated when biotinylated human transferrin was used instead of biotinylated porcine transferrin. The 64 kDa polypeptide was the more easily removed from Sepharose-coupled porcine transferrin and only the 99 kDa polypeptide appeared to be an outer-membrane protein. While these results suggest that the 99 kDa polypeptide represents the porcine transferrin receptor of A. pleuropneumoniae, and that the 64 kDa polypeptide represents an associated protein serving an accessory role, other interpretations are also possible.
Ricard
M A
MA
Department of Microbiology, Macdonald Campus of McGill University, Ste Anne de Bellevue, Québec, Canada.
Archibald
F S
FS
Niven
D F
DF
D016977:Actinobacillus pleuropneumoniae / Q000302:isolation & purification*; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D001426:Bacterial Proteins; D001710:Biotin; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D033862:Iron-Binding Proteins; D033902:Periplasmic Binding Proteins; D011990:Receptors, Transferrin / Q000302:isolation & purification* / Q000378:metabolism; D013552:Swine; D014168:Transferrin / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D033862:Iron-Binding Proteins; D033902:Periplasmic Binding Proteins; D011990:Receptors, Transferrin; D014168:Transferrin; D001710:Biotin