Isolation and characterization of fimbriae from a sparsely fimbriated strain of Porphyromonas gingivalis.

Applied and Environmental Microbiology
H SojarR Genco

Abstract

Porphyromonas gingivalis W50 (ATCC 53978) possesses the gene for fimbriae; however, the surface-expressed fimbriae are sparse and have not been previously isolated and characterized. We purified fimbriae from strain W50 to homogeneity by ammonium sulfate precipitation and reverse-phase high-performance liquid chromatography [H. T. Sojar, N. Hamada, and R. J. Genco, Protein Expr. Purif. 9(1):49-52, 1997]. Negative staining of purified fimbriae viewed by electron microscopy revealed that the fimbriae were identical in diameter to fimbriae of other P. gingivalis strains, such as 2561, but were shorter in length. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, the apparent molecular weight of isolated fimbrillin from strain W50 was found to be identical to that of the fimbrillin molecule of strain 2561. Unlike 2561 fimbriae, W50 fimbriae, under reducing condition, exhibited a monomeric structure on SDS-PAGE at room temperature. However, under nonreduced conditions, even at 100 degrees C, no monomer was observed. In immunoblot analysis as well as immunogold labeling of isolated fimbriae, polyclonal antibodies against 2561 fimbriae, as well as antibodies against peptide I (V-V-M-A-N-T-G-A-M-E-V-G-K-T...Continue Reading

References

Oct 1, 1979·Journal of Clinical Periodontology·A C TannerS S Socransky
Apr 1, 1977·Journal of General Microbiology·J A MorrisW J Sojka
Oct 1, 1979·Journal of Clinical Periodontology·J Slots
Mar 15, 1991·Biochemical and Biophysical Research Communications·H T SojarR J Genco
Dec 1, 1991·Infection and Immunity·A M De NardinR J Genco
Apr 1, 1988·Journal of Clinical Periodontology·A D HaffajeeD J Smith
Aug 1, 1995·Protein Expression and Purification·J Y LeeR J Genco
Feb 1, 1997·Protein Expression and Purification·H T SojarR J Genco

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Citations

Jan 13, 2001·Oral Microbiology and Immunology·P J BakerD C Roopenian
May 29, 2002·Oral Microbiology and Immunology·N SlakeskiE C Reynolds
Oct 16, 1999·Periodontology 2000·S C HoltC A Genco
Mar 21, 2012·FEMS Immunology and Medical Microbiology·Young-Suk ChoiJin-Yong Lee
Feb 21, 2003·Journal of Periodontology·Howard K KuramitsuMinshan Qi

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