FEMS Microbiology Letters · 2001 · 138 citations · 21 references
DentistryMicrobiologyOral BiologyBacteriologyMicrobial EcologyOral MicrobiologyBacterial BiofilmsBiofilm FormationDental CariesDental PlaqueMicrobial BiofilmsMicrobiology Of Dental CariesAmino AcidsDental Plaque BiofilmsDental Plaque MetabolismMass Spectrometry ProteomicsPh DependenceProtein Expression LevelsTwo-dimensional ElectrophoresisTwo-dimensional Gel Electrophoresis
Streptococcus mutans, a major causal agent of dental caries, functions in nature as a component of a biofilm on teeth (dental plaque) and yet very little information is available on the physiology of the organism in such surface-associated communities. As a consequence, we undertook to examine the synthesis of proteins by planktonic and biofilm cells growing in a biofilm chemostat at pH 7.5 at a dilution rate of 0.1 h(-1) (mean generation time=7 h). Cells were incubated with (14)C-labelled amino acids, the proteins extracted and separated by two-dimensional electrophoresis followed by autoradiography and computer-assisted image analysis. Of 694 proteins analysed, 57 proteins were enhanced 1.3-fold or greater in biofilm cells compared to planktonic cells with 13 only expressed in sessile cells. Diminished protein expression was observed with 78 proteins, nine of which were not expressed in biofilm cells. The identification of enhanced and diminished proteins by mass spectrometry and computer-assisted protein sequence analysis revealed that, in general, glycolytic enzymes involved in acid formation were repressed in biofilm cells, while biosynthetic processes were enhanced. The results show that biofilm cells possess novel proteins, of as yet unknown function, that are not present in planktonic cells.
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Mass Spectrometric Sequencing of Proteins from Silver-Stained Polyacrylamide Gels
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