Abstract
Proteomic analyses are increasingly implemented to investigate the particular physiology of (naturally or artificially) immobilized microorganisms. The protein maps of immobilized cells are compared with those of suspended counterparts to reveal alterations in protein expression induced by the microbial mode of growth. Proteins whose amount significantly varies between free and immobilized cells are identified by mass spectrometry and referring the peptide fingerprints to published databases. This proteomic approach is illustrated here using Pseudomonas aeruginosa cells grown as biofilms on clay beads for 24 or 48 h. Both the growth mode (suspended or attached) and the incubation time were shown to control the expression level of a large number of proteins by P. aeruginosa. Proteins whose amount significantly varied in biofilm organisms compared to suspended bacteria could be divided into three main classes, namely proteins linked to metabolic processes, proteins involved in adaptation and protection, and membrane proteins.
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References
Costerton J. W., Lewandowski Z., Caldwell D. E., Korber D. R. and Lappin-Scott H. M. (1995) Microbial biofilms. Annu. Rev. Microbiol. 49, 711–745.
Davies D. (2003) Understanding biofilm resistance to antibacterial agents. Nature Rev. 2, 114–122.
Fleischmann R. D., Adams M. D., White O., et al. (1995) Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science 269, 496–521.
Wilkins M. R., Sanchez J. C., Gooley A. A., Appel R. D., Humphery-Smith I., Hochstrasser D. F., and Williams K. L. (1995) Progress in proteome projects: why all proteins expressed by a genome should be identified and how to do it. Biotechnol. Genet. Eng. Rev. 13, 19–50.
Sauer K. (2003) The genomics and proteomics of biofilm formation. Genome Biol. 4, 219.
Vilain S., Cosette P., Zimmerlin I., Dupont J.-P., Junter G.-A., and Jouenne T. (2004) The biofilm proteome: homogeneity or versatility? J. Proteome Res. 3, 132–136.
Caldwell D. E. (1995) Cultivation and study of biofilm communities. In: Microbial Biofilms, Lappin-Sott H. M. and J.W. Costerton, eds., Cambridge University Press, Cambridge, UK, pp. 64–79.
Brown M. R. W. and Barker J. (1999) Unexplored reservoirs of pathogenic bacteria: protozoa and biofilms. Trends Microbiol. 7, 46–50.
Schembri M. A. and Klemm P. (2001) Coordinate gene regulation by fimbriaeinduced signal transduction. EMBO J. 20, 3074–3081.
Otto K. and Silahvy T. J. (2002) Surface sensing and adhesion of Escherichia coli under control of the Cpx signalling pathway. Proc. Nat. Acad. Sci. USA 99, 2287–2292.
Aires J. R., Köhler T., Nikaido H., and Plésiat P. (1999) Involvement of an active efflux system in the natural resistance of Pseudomonas aeruginosa to aminoglycosides. Antimicrob. Agents Chemother. 43, 2624–2628.
Köhler T., Michea-Hamzehpour M., Epp S. F., and Pechere J. C. (1999) Carbapenem activities against Pseudomonas aeruginosa: respective contributions of OprD and efflux systems. Antimicrob. Agents Chemother. 43, 424–427.
Sauer K. and Camper A. K. (2001) Characterization of phenotypic changes in Pseudomonas putida in response to surface-associated growth. J. Bacteriol. 183, 6579–6589.
Svensäter G., Welin J., Wilkins J. C., Beighton D., and Hamilton I. R. (2001) Protein expression by planktonic and biofilm cells of Streptococcus mutans. FEMS Microbiol. Lett. 205, 139–146.
Sauer K., Camper A. K., Erlich G. A., Costerton J. W., and Davies D. G. (2002) Pseudomonas aeruginosa displays multiple phenotypes during development as a biofilm. J. Bacteriol. 184, 1140–1154.
Schembri M. A., Kjærgaard K., and Klemm P. (2003) Global gene expression in Escherichia coli biofilms. Mol. Microbiol. 48, 253–267.
Vilain S., Cosette P., Hubert M., Lange C., Junter G.-A., and Jouenne T. (2004) Comparative proteomic analysis of planktonic and immobilized Pseudomonas aeruginosa cells: a multivariate statistical approach. Anal. Biochem. 329, 120–130.
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Coquet, L., Vilain, S., Cosette, P., Jouenne, T., Junter, GA. (2006). A Proteomic Approach to Biofilm Cell Physiology. In: Guisan, J.M. (eds) Immobilization of Enzymes and Cells. Methods in Biotechnologyâ„¢, vol 22. Humana Press. https://doi.org/10.1007/978-1-59745-053-9_35
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DOI: https://doi.org/10.1007/978-1-59745-053-9_35
Publisher Name: Humana Press
Print ISBN: 978-1-58829-290-2
Online ISBN: 978-1-59745-053-9
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