title: Purification, characterization and immunolocalization of fimbrial protein from Porphyromonas (bacteroides) gingivalis.
abstract: Rapid and reproducible method is described here for the purification of the 43 kDa fimbrial protein from P. gingivalis by preferential fractionation in the presence of 1% SDS and 0.2M of a bivalent cation at pH 6.5. Homogeneity of the purified 43 kDa was confirmed by SDS-PAGE and Western blot analysis using monoclonal and polyclonal antibodies raised against this protein. Amino acid composition and the amino acid sequence of the first 30 amino acid residues of the purified fimbriae are consistent with the composition and sequence predicted from the cloned gene of the fimbrial subunit. Circular dichroism spectra shows high levels of beta-sheet structure. The purified 43 kDa polymer shows fimbriae-like morphology under the electron microscope. Ultrastructural localization of the 43 kDa protein by the immunogold technique revealed specific labeling of the fimbriae with a diameter of approximately 3.5 to 5.0 nm. Localization of this protein suggest that the 43 kDa component is a fimbrial subunit.
authors.affiliation: Department of Oral Biology, State University of New York, Buffalo, Buffalo 14214.
authors.forename: H T
authors.initials: HT
authors.lastname: Sojar
authors.forename: J Y
authors.initials: JY
authors.lastname: Lee
authors.forename: G S
authors.initials: GS
authors.lastname: Bedi
authors.forename: M I
authors.initials: MI
authors.lastname: Cho
authors.forename: R J
authors.initials: RJ
authors.lastname: Genco
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D001439:Bacteroides / Q000032:analysis* / Q000276:immunology; D002942:Circular Dichroism; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000737:chemistry*; D007150:Immunohistochemistry; D008970:Molecular Weight
chemical_list: D001426:Bacterial Proteins; C063055:fimbrillin; D036781:Fimbriae Proteins