PMID: 8941761Apr 1, 1996Paper

Proteolytic artifacts in SDS-PAGE analysis of selected periodontal pathogens

Oral Microbiology and Immunology
M F WeidnerD Mayrand

Abstract

The aim of the study was to examine whether proteolytic artifacts, which result in a loss and poor resolution of protein bands, occur during sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis analysis of cellular proteins from selected proteolytic (Porphyromonas gingivalis, Prevotella nigrescens and Treponema denticola) and non-proteolytic (Fusobacterium nucleatum) bacteria. Conditions to limit or prevent proteolysis were also investigated. Bacterial cells were incubated in solubilizing buffer (SDS+ beta mercaptoethanol) at room temperature for various periods of time before boiling. A control assay consisted of trichloroacetic acid-treated bacterial cells. Cellular proteins were separated by electrophoresis and stained with Coomassie blue. Proteolysis occurred very rapidly in the case of P. gingivalis (< 30 s), whereas a longer incubation time (> 1 h) was required to observe similar effects in P. nigrescens and T. denticola. No proteolysis was observed for F. nucleatum. In all cases, heat (100 degrees C) and low pH (< 4) treatments of bacterial cells could avoid production of proteolytic artifacts. Incorporation of specific protease inhibitors before solubilization of bacteria could also prevent proteolysis. More ...Continue Reading

References

Jan 1, 1991·Critical Reviews in Oral Biology and Medicine : an Official Publication of the American Association of Oral Biologists·S C Holt, T E Bramanti
Jan 1, 1991·International Archives of Allergy and Applied Immunology·D M Kemeny, D Diaz-Sanchez
Mar 1, 1985·Canadian Journal of Microbiology·G D Williams, S C Holt
Oct 1, 1988·Infection and Immunity·V J UittoB C McBride
Dec 1, 1980·Journal of Clinical Microbiology·S B Leschine, E Canale-Parola
Oct 1, 1993·Comparative Biochemistry and Physiology. Comparative Physiology·M S RendellD A Finney
Oct 1, 1993·Comparative Biochemistry and Physiology. Comparative Physiology·M O Nielsen, K Jakobsen
Oct 1, 1993·Comparative Biochemistry and Physiology. C, Comparative Pharmacology and Toxicology·G KochR J Walker
Nov 1, 1993·Journal of Bacteriology·T E Bramanti, S C Holt
Jan 1, 1960·Annual Review of Biochemistry·B S HARTLEY

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Citations

Nov 24, 1999·FEMS Microbiology Letters·D E LopatinC E Shelburne
Oct 24, 1997·Nature·E H Davidson
Jan 1, 1997·Critical Reviews in Oral Biology and Medicine : an Official Publication of the American Association of Oral Biologists·D H MeyerP M Fives-Taylor

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