Mv. Marandi et Kr. Mittal, CHARACTERIZATION OF AN OUTER-MEMBRANE PROTEIN OF PASTEURELLA-MULTOCIDA BELONGING TO THE OMPA FAMILY, Veterinary microbiology, 53(3-4), 1996, pp. 303-314
The outer membrane vesicle and N-lauroylsarcosine-insoluble protein pr
eparations of Pasteurella multocida 656 were analyzed by sodium dodecy
l sulfate-polyacrylamide gel electrophoresis. A major outer membrane p
rotein (OMP) was found to be heat-modifiable, having a molecular mass
of 28 kDa when the OMP preparation was solubilized at 60 degrees C and
a molecular mass of 37 kDa when it was solubilized at 100 degrees C.
A monoclonal antibody, designated mAb MT4.1, was generated against hea
t-modifiable OMP of P. multocida. This mAb reacted with the heat-modif
iable OMP irrespective of the temperature at which it was solubilized,
as demonstrated by immunoblot results. The heat-modifiable OMP of P.
multocida showed a significant N-terminal amino acid sequence homology
with OmpA family. Immunoelectron microscopic study revealed that the
mAb MT4.1 epitope was not surface exposed on the intact bacterium. The
mAb MT4.1 reacted with all the reference strains of 5 capsular and 16
somatic serotypes, as well as with 75 field strains of P. multocida i
n immunoblot assay. This mAb MT4.1 also reacted with strains of variou
s other Pasteurella species such as P. stomatis, P. aerogenes, P. gall
inarum, P. betti, P. sp. B, P. SP-g and P. canis, but not with strains
of 12 other Gram-negative bacteria. These results indicated that this
protein carried a genus-specific epitope and mAb MT4.1 may be useful
for identification of Pasteurella species. This is the first report in
which a major heat-modifiable OMP has been identified and characteriz
ed using a mAb, and has been shown belonging to the OmpA family.