Isolation and characterization of Pseudomonas pseudomallei flagellin proteins.
Flagellin proteins from several different strains of Pseudomonas pseudomallei have been isolated and purified to homogeneity by mechanical shearing and differential centrifugation techniques. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis yielded flagellin monomer protein bands with an estimated M(r) of 43,400. No lipopolysaccharide contamination of the purified protein preparations was detectable by silver staining of flagellin displayed on polyacrylamide gels and by Western immunoblotting with P. pseudomallei antilipopolysaccharide monoclonal antibody. NH2-terminal amino acid sequence analysis of the flagellin protein of P. pseudomallei 319a revealed significant homology with flagellins from Proteus mirabilis, Bordetella bronchiseptica, and Pseudomonas aeruginosa PAK. Rabbit polyclonal antiserum raised against the 319a flagellin protein reacted with 64 of 65 P. pseudomallei strains tested. The polyclonal antiserum proved effective in inhibiting the motility of these organisms in motility agar plates. Passive immunization studies demonstrated that 319a flagellin-specific antiserum was capable of protecting diabetic rats from challenge with a heterologous P. pseudomallei strain.
Brett
P J
PJ
Department of Microbiology and Infectious Diseases, University of Calgary Health Sciences Centre, Alberta, Canada.
Mah
D C
DC
Woods
D E
DE
D000595:Amino Acid Sequence; D000818:Animals; D016957:Burkholderia pseudomallei / Q000737:chemistry / Q000276:immunology*; D005408:Flagellin / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D007116:Immunization, Passive; D008070:Lipopolysaccharides / Q000032:analysis; D008297:Male; D008554:Melioidosis / Q000517:prevention & control; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley
D008070:Lipopolysaccharides; D005408:Flagellin