Comparison of S. aureus Proteomic Profiles from Biofilm and Planktonic Growth Conditions using 2D- Gel Electrophoresis.
Bacteria growing as biofilms are distinct from the same bacteria growing as planktonic cells. Biofilms cells show increased resistance to antimicrobial, immunological, predatory, and chemical attack than planktonic cells. Most studies on bacterial diseases use planktonic bacteria. The objective of this study was to identify expressed proteins that are unique to Staphylococcus aureus biofilm mode of growth. S. aureus was grown in tryptic soy broth and Dulbecco's modified eagle medium at biofilm and planktonic growth conditions. Protein samples were cleaned up and separated according to their electrophoretic mobility using 7 cm IPG strips (pH 3–10 and pH 4–7) on 2D gel electrophoresis. Expressed proteins of both growth conditions were compared. Data analysis revealed that the expression of S. aureus proteins from planktonic and biofilm was higher in TSB media than DMEM media. Biofilm growth condition showed higher intensity of expressed proteins and new expressed proteins were observed. One protein was found to be upregulated in planktonic growth condition. Additionally, the majority of the proteins were clustered in the area of acidic region (pH 4–7). 2D-gel electrophoresis is a powerful and widely used method for the proteomic analysis. Biofilms represent a realistic representation of bacterial behavior and organisms are capable of altering their physiology in the surrounded environments. The results could help to illustrate the differences in pathogenesis between biofilm and planktonic cells in any model of disease. This will identify biological markers to improve the diagnostics, treatment, and prevention of S. aureus biofilms.
Beenken, K.E.; Dunman, P.M.; McAleese, F.; Macapagal, D.; Murphy, E.; Projan, S.J., and Smeltzer, M.S. (2004) Global gene expression in Staphylococcus aureus biofilms. J. Bacteriol., 186: 4665-4684.
Brady, R.A.; Leid, J.G.; Camper, A.K.; Costerton, J.W., and Shirtliff, M.E. (2006) Identification of Staphylococcus aureus proteins recognized by the antibody-mediated immune response to a biofilm infection. Infect. Immun.,74: 3415-3426.
Branda, S.S.; Vik, S.; Friedman, L., and Kolter, R. (2005) Biofilms: the matrix revisited. Trends Microbiol., 13: 20-26.
Cheung, A.L.; Bayer, A.S.; Zhang, G.; Gresham, H., and Xiong, Y.Q. (2004) Regulation of virulence determinants in vitro and in vivo in Staphylococcus aureus. FEMS. Immunol. Med. Microbiol., 40: 1-9.
Cordwell, S.J.; Larsen, M.R.; Cole, R.T., and Walsh, B.J. (2002) Comparative proteomics of Staphylococcus aureus and the response of methicillin-resistant and methicillin-sensitive strains to Triton X-100. Microbiology, 148: 2765-2781.
Delaune, A.; Dubrac, S.; Blanchet, C.; Poupel, O.; Mader, U.; Hiron, A., and Msadek, T. (2012) The WalKR system controls major staphylococcal virulence genes and is involved in triggering the host inflammatory response. Infect. Immun., 80: 3438-3453.
Flemming, H.C., and Wingender, J. (2010) The biofilm matrix. Nat. Rev. Microbiol., 8: 623-633.
Foster, T.J. (2005) Immune evasion by staphylococci. Nat. Rev. Microbiol., 3: 948-958.
Gjodsbol, K.; Christensen, J.J.; Karlsmark, T.; Jorgensen, B.; Klein, B.M., and Krogfelt, K.A. (2006) Multiple bacterial species reside in chronic wounds: a longitudinal study. Int. Wound. J., 3: 225-231.
Hogt, A.H.; Dankert, J., and Feijen, J. (1983) Encapsulation, slime production and surface hydrophobicity of coagulase-negative staphylococci. FEMS. Microbiology Letters, 18: 211-215.
James, G.A.; Swogger, E.; Wolcott, R.; Pulcini, E.; Secor, P.; Sestrich, J., and Stewart, P.S. (2008) Biofilms in Chronic Wounds. Wound. Rep. Reg., 16: 37-44.
Kirker, K.R.; James, G.A.; Fleckman, P.; Olerud, J.E., and Stewart, P.S. (2012) Differential effects of planktonic and biofilm MRSA on human fibroblasts. Wound Repair Regen., 20: 253-261.
Kristian, S.A.; Golda, T.; Ferracin, F.; Cramton, S.E.; Neumeister, B.; Peschel, A., and Landmann, R. (2004) The ability of biofilm formation does not influence virulence of Staphylococcus aureus and host response in a mouse tissue cage infection model. Microb. Pathog., 36: 237-245.
Matsumoto, H.; Haniu, H.; Kurien, B.T., and Komori, N. (2012) Two-dimensional gel electrophoresis: glass tube-based IEF followed by SDS-PAGE. Methods Mol. Biol., 869: 267-273.
Oogai, Y.; Matsuo, M.; Hashimoto, M.; Kato, F.; Sugai, M., and Komatsuzawa, H. (2011) Expression of virulence factors by Staphylococcus aureus grown in serum. Appl. Environ. Microbiol., 77: 8097-8105.
Otto, M. (2008) Staphylococcal biofilms. Curr. Top. Microbiol. Immunol., 322: 207-228.
Resch, A.; Rosenstein, R.; Nerz, C., and Gotz, F. (2005) Differential gene expression profiling of Staphylococcus aureus cultivated under biofilm and planktonic conditions. Appl. Environ. Microbiol., 71: 2663-2676.
Rosen, R.; Becher, D.; Buttner, K.; Biran, D.; Hecker, M., and Ron, E.Z. (2004) Highly phosphorylated bacterial proteins. Proteomics, 4: 3068-3077.
Sauer, K.; Camper, A.K.; Ehrlich, G.D.; Costerton, J.W., and Davies, D.G. (2002) Pseudomonas aeruginosa displays multiple phenotypes during development as a biofilm. J. Bacteriol., 184: 1140-1154.
Schierle, C.F.; De la Garza, M.; Mustoe, T.A., and Galiano, R.D. (2009) Staphylococcal biofilms impair wound healing by delaying reepithelialization in a murine cutaneous wound model. Wound Repair Regen., 17: 354-359.
Secor, P.R.; James, G.A.; Fleckman, P.; Olerud, J.E.; McInnerney, K., and Stewart, P.S. (2011) Staphylococcus aureus Biofilm and Planktonic cultures differentially impact gene expression, mapk phosphorylation, and cytokine production in human keratinocytes. BMC Microbiol 11: 143.
Stewart, P.S., and Costerton, J.W. (2001) Antibiotic resistance of bacteria in biofilms. Lancet, 358: 135-138.
Stoodley, P.; Sauer, K.; Davies, D.G., and Costerton, J.W. (2002) Biofilms as complex differentiated communities. Annu. Rev. Microbiol., 56: 187-209.
Tankersley, A.; Frank, M.B.; Bebak, M., and Brennan, R. (2014) Early effects of Staphylococcus aureus biofilm secreted products on inflammatory responses of human epithelial keratinocytes. J. Inflamm., 11: 17.
Valle, J.; Toledo-Arana, A.; Berasain, C.; Ghigo, J.M.; Amorena, B.; Penadés, J.R., and Lasa, I. (2003) SarA and not sigmaB is essential for biofilm development by Staphylococcus aureus. Mol. Microbiol., 48: 1075-1087.
You, S.A., and Wang, Q.K. (2006) Proteomics with two-dimensional gel electrophoresis and mass spectrometry analysis in cardiovascular research. Methods Mol. Med., 129: 15-26.
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