Characterization of two lipoproteins in Pasteurella multocida.
An in vivo expression technology (IVET) system was previously developed and used to identify Pasteurella multocida genes, which are upregulated during infection of the host. Of the many genes identified, two encoded products which showed similarity to the Haemophilus influenzae lipoproteins, protein D and PCP, which have been shown to stimulate heterologous immunity against infection with H. influenzae. Therefore, the lipoprotein homologues in P. multocida, designated GlpQ and PCP, were investigated. GlpQ and PCP were shown to be lipoproteins by demonstrating that post-translational processing of the proteins was inhibited by globomycin. The P. multocida GlpQ homologue showed glycerophosphodiester phosphodiesterase enzyme activity, indicating that it is a functional homologue of other characterized GlpQ enzymes. Using surface immunoprecipitation, PCP was found to be surface exposed, but GlpQ was not. Non-lipidated forms of GlpQ and PCP were expressed and purified from Escherichia coli and used to vaccinate mice. However, mice were not protected from challenge with live P. multocida. The lipoproteins were then expressed in E. coli in the lipidated form and used to vaccinate mice and chickens. Protection against challenge with live P. multocida was not observed.
Lo
Miranda
M
Bacterial Pathogenesis Research Group, Department of Microbiology, Monash University, Melbourne,. 3800, Vic, Australia.
Boyce
John D
JD
Wilkie
Ian W
IW
Adler
Ben
B
D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D001428:Bacterial Vaccines / Q000008:administration & dosage* / Q000276:immunology*; D015153:Blotting, Western; D002645:Chickens; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D008074:Lipoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D006899:Mixed Function Oxygenases / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D010325:Pasteurella / Q000235:genetics / Q000276:immunology*; D010326:Pasteurella Infections / Q000276:immunology* / Q000517:prevention & control*; D010727:Phosphoric Diester Hydrolases / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis; D014611:Vaccination; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000276:immunology
D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D008074:Lipoproteins; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D006899:Mixed Function Oxygenases; C069781:pentachlorophenol monooxygenase; D010727:Phosphoric Diester Hydrolases; C087921:glpQ protein, Bacteria