Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1995 Aug;61(8):2863–2872. doi: 10.1128/aem.61.8.2863-2872.1995

Two-Dimensional Gel Electrophoresis Analysis of the Response of Pseudomonas putida KT2442 to 2-Chlorophenol

C G Lupi, T Colangelo, C A Mason
PMCID: PMC1388547  PMID: 16535093

Abstract

The effects of exposure of Pseudomonas putida KT2442 to 2-chlorophenol as a model for the chemical stress response were examined by two-dimensional polyacrylamide gel electrophoresis. Individual protein concentrations were determined at 45, 65, and 95 min following the addition of 2-chlorophenol at a concentration of 1.63 mM to exponentially growing cultures of P. putida KT2442 by silver staining the separated proteins. The changes in the protein concentrations could be classified into four categories, namely those which increased continuously during exposure, those which decreased in concentration, those which showed a concentration peak at some point following exposure, and those which were essentially unaffected. Thirty proteins with isoelectric points between pH 4 and 6 increased in concentration, 27 decreased, and 90 had a concentration maximum or minimum between 45 and 95 min. Of those proteins with isoelectric points between 5.5 and 10, 68 increased in concentration, 39 decreased in concentration, and 47 showed a concentration peak in the middle of the sampling period. Thus, in the evaluation of the stress response, a functional description requires an understanding both of proteins which are required at higher concentrations and of those whose presence appears to be no longer essential.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Appel R. D., Hochstrasser D. F., Funk M., Vargas J. R., Pellegrini C., Muller A. F., Scherrer J. R. The MELANIE project: from a biopsy to automatic protein map interpretation by computer. Electrophoresis. 1991 Oct;12(10):722–735. doi: 10.1002/elps.1150121006. [DOI] [PubMed] [Google Scholar]
  2. Bjellqvist B., Pasquali C., Ravier F., Sanchez J. C., Hochstrasser D. A nonlinear wide-range immobilized pH gradient for two-dimensional electrophoresis and its definition in a relevant pH scale. Electrophoresis. 1993 Dec;14(12):1357–1365. doi: 10.1002/elps.11501401209. [DOI] [PubMed] [Google Scholar]
  3. Blom A., Harder W., Matin A. Unique and overlapping pollutant stress proteins of Escherichia coli. Appl Environ Microbiol. 1992 Jan;58(1):331–334. doi: 10.1128/aem.58.1.331-334.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Faber F., Egli T., Harder W. Transient repression of the synthesis of OmpF and aspartate transcarbamoylase in Escherichia coli K12 as a response to pollutant stress. FEMS Microbiol Lett. 1993 Aug 1;111(2-3):189–195. doi: 10.1111/j.1574-6968.1993.tb06384.x. [DOI] [PubMed] [Google Scholar]
  5. Gage D. J., Neidhardt F. C. Adaptation of Escherichia coli to the uncoupler of oxidative phosphorylation 2,4-dinitrophenol. J Bacteriol. 1993 Nov;175(21):7105–7108. doi: 10.1128/jb.175.21.7105-7108.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Georgopoulos C., Welch W. J. Role of the major heat shock proteins as molecular chaperones. Annu Rev Cell Biol. 1993;9:601–634. doi: 10.1146/annurev.cb.09.110193.003125. [DOI] [PubMed] [Google Scholar]
  7. Givskov M., Eberl L., Molin S. Responses to nutrient starvation in Pseudomonas putida KT2442: two-dimensional electrophoretic analysis of starvation- and stress-induced proteins. J Bacteriol. 1994 Aug;176(16):4816–4824. doi: 10.1128/jb.176.16.4816-4824.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Görg A., Postel W., Günther S. The current state of two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis. 1988 Sep;9(9):531–546. doi: 10.1002/elps.1150090913. [DOI] [PubMed] [Google Scholar]
  9. Görg A. Two-dimensional electrophoresis with immobilized pH gradients: current state. Biochem Soc Trans. 1993 Feb;21(1):130–132. doi: 10.1042/bst0210130. [DOI] [PubMed] [Google Scholar]
  10. Heitzer A., Mason C. A., Hamer G. Heat shock gene expression in continuous cultures of Escherichia coli. J Biotechnol. 1992 Jan;22(1-2):153–169. doi: 10.1016/0168-1656(92)90139-z. [DOI] [PubMed] [Google Scholar]
  11. Herendeen S. L., VanBogelen R. A., Neidhardt F. C. Levels of major proteins of Escherichia coli during growth at different temperatures. J Bacteriol. 1979 Jul;139(1):185–194. doi: 10.1128/jb.139.1.185-194.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kalb V. F., Jr, Bernlohr R. W. A new spectrophotometric assay for protein in cell extracts. Anal Biochem. 1977 Oct;82(2):362–371. doi: 10.1016/0003-2697(77)90173-7. [DOI] [PubMed] [Google Scholar]
  13. Kertesz M. A., Leisinger T., Cook A. M. Proteins induced by sulfate limitation in Escherichia coli, Pseudomonas putida, or Staphylococcus aureus. J Bacteriol. 1993 Feb;175(4):1187–1190. doi: 10.1128/jb.175.4.1187-1190.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kjelleberg S., Albertson N., Flärdh K., Holmquist L., Jouper-Jaan A., Marouga R., Ostling J., Svenblad B., Weichart D. How do non-differentiating bacteria adapt to starvation? Antonie Van Leeuwenhoek. 1993;63(3-4):333–341. doi: 10.1007/BF00871228. [DOI] [PubMed] [Google Scholar]
  15. Kroh H. E., Simon L. D. The ClpP component of Clp protease is the sigma 32-dependent heat shock protein F21.5. J Bacteriol. 1990 Oct;172(10):6026–6034. doi: 10.1128/jb.172.10.6026-6034.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lange R., Hengge-Aronis R. Identification of a central regulator of stationary-phase gene expression in Escherichia coli. Mol Microbiol. 1991 Jan;5(1):49–59. doi: 10.1111/j.1365-2958.1991.tb01825.x. [DOI] [PubMed] [Google Scholar]
  17. Lemaux P. G., Herendeen S. L., Bloch P. L., Neidhardt F. C. Transient rates of synthesis of individual polypeptides in E. coli following temperature shifts. Cell. 1978 Mar;13(3):427–434. doi: 10.1016/0092-8674(78)90317-3. [DOI] [PubMed] [Google Scholar]
  18. Nyström T., Neidhardt F. C. Cloning, mapping and nucleotide sequencing of a gene encoding a universal stress protein in Escherichia coli. Mol Microbiol. 1992 Nov;6(21):3187–3198. doi: 10.1111/j.1365-2958.1992.tb01774.x. [DOI] [PubMed] [Google Scholar]
  19. Nyström T., Neidhardt F. C. Isolation and properties of a mutant of Escherichia coli with an insertional inactivation of the uspA gene, which encodes a universal stress protein. J Bacteriol. 1993 Jul;175(13):3949–3956. doi: 10.1128/jb.175.13.3949-3956.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nyström T., Olsson R. M., Kjelleberg S. Survival, stress resistance, and alterations in protein expression in the marine vibrio sp. strain S14 during starvation for different individual nutrients. Appl Environ Microbiol. 1992 Jan;58(1):55–65. doi: 10.1128/aem.58.1.55-65.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Odberg-Ferragut C., Espigares M., Dive D. Stress protein synthesis, a potential toxicity marker in Escherichia coli. Ecotoxicol Environ Saf. 1991 Jun;21(3):275–282. doi: 10.1016/0147-6513(91)90066-x. [DOI] [PubMed] [Google Scholar]
  22. Spector M. P., Aliabadi Z., Gonzalez T., Foster J. W. Global control in Salmonella typhimurium: two-dimensional electrophoretic analysis of starvation-, anaerobiosis-, and heat shock-inducible proteins. J Bacteriol. 1986 Oct;168(1):420–424. doi: 10.1128/jb.168.1.420-424.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. VanBogelen R. A., Kelley P. M., Neidhardt F. C. Differential induction of heat shock, SOS, and oxidation stress regulons and accumulation of nucleotides in Escherichia coli. J Bacteriol. 1987 Jan;169(1):26–32. doi: 10.1128/jb.169.1.26-32.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. VanBogelen R. A., Sankar P., Clark R. L., Bogan J. A., Neidhardt F. C. The gene-protein database of Escherichia coli: edition 5. Electrophoresis. 1992 Dec;13(12):1014–1054. doi: 10.1002/elps.11501301203. [DOI] [PubMed] [Google Scholar]
  25. Völker U., Engelmann S., Maul B., Riethdorf S., Völker A., Schmid R., Mach H., Hecker M. Analysis of the induction of general stress proteins of Bacillus subtilis. Microbiology. 1994 Apr;140(Pt 4):741–752. doi: 10.1099/00221287-140-4-741. [DOI] [PubMed] [Google Scholar]
  26. Welch T. J., Farewell A., Neidhardt F. C., Bartlett D. H. Stress response of Escherichia coli to elevated hydrostatic pressure. J Bacteriol. 1993 Nov;175(22):7170–7177. doi: 10.1128/jb.175.22.7170-7177.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yura T., Nagai H., Mori H. Regulation of the heat-shock response in bacteria. Annu Rev Microbiol. 1993;47:321–350. doi: 10.1146/annurev.mi.47.100193.001541. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES