This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Elowitz, M. B.
Right arrow Articles by Leibler, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Elowitz, M. B.
Right arrow Articles by Leibler, S.

 Previous Article  |  Next Article 

Journal of Bacteriology, January 1999, p. 197-203, Vol. 181, No. 1
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Protein Mobility in the Cytoplasm of Escherichia coli

Michael B. Elowitz,1,2,* Michael G. Surette,2,dagger Pierre-Etienne Wolf,1,2,Dagger Jeffry B. Stock,2 and Stanislas Leibler1,2

Departments of Physics1 and Molecular Biology,2 Princeton University, Princeton, New Jersey 08544

Received 7 August 1998/Accepted 21 October 1998

The rate of protein diffusion in bacterial cytoplasm may constrain a variety of cellular functions and limit the rates of many biochemical reactions in vivo. In this paper, we report noninvasive measurements of the apparent diffusion coefficient of green fluorescent protein (GFP) in the cytoplasm of Escherichia coli. These measurements were made in two ways: by photobleaching of GFP fluorescence and by photoactivation of a red-emitting fluorescent state of GFP (M. B. Elowitz, M. G. Surette, P. E. Wolf, J. Stock, and S. Leibler, Curr. Biol. 7:809-812, 1997). The apparent diffusion coefficient, Da, of GFP in E. coli DH5alpha was found to be 7.7 ± 2.5 µm2/s. A 72-kDa fusion protein composed of GFP and a cytoplasmically localized maltose binding protein domain moves more slowly, with Da of 2.5 ± 0.6 µm2/s. In addition, GFP mobility can depend strongly on at least two factors: first, Da is reduced to 3.6 ± 0.7 µm2/s at high levels of GFP expression; second, the addition to GFP of a small tag consisting of six histidine residues reduces Da to 4.0 ± 2.0 µm2/s. Thus, a single effective cytoplasmic viscosity cannot explain all values of Da reported here. These measurements have implications for the understanding of intracellular biochemical networks.

* Corresponding author. Mailing address: Lewis Thomas Lab, Washington Rd., Princeton, NJ 08544. Phone: (609) 258-1574. Fax: (609) 258-6175. E-mail: melowitz{at}princeton.edu.

dagger Present address: Department of Microbiology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada T2N 4N1.

Dagger Present address: Centre de Recherches sur les Très Basses Températures, CNRS, Laboratoire associé à l'Université Joseph Fourier, F-38042 Grenoble Cedex 9, France.

Journal of Bacteriology, January 1999, p. 197-203, Vol. 181, No. 1
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Spitzer, J., Poolman, B. (2009). The Role of Biomacromolecular Crowding, Ionic Strength, and Physicochemical Gradients in the Complexities of Life's Emergence. Microbiol. Mol. Biol. Rev. 73: 371-388 [Abstract] [Full Text]  
  • Konopka, M. C., Sochacki, K. A., Bratton, B. P., Shkel, I. A., Record, M. T., Weisshaar, J. C. (2009). Cytoplasmic Protein Mobility in Osmotically Stressed Escherichia coli. J. Bacteriol. 191: 231-237 [Abstract] [Full Text]  
  • Gregory, J. A., Becker, E. C., Pogliano, K. (2008). Bacillus subtilis MinC destabilizes FtsZ-rings at new cell poles and contributes to the timing of cell division. Genes Dev. 22: 3475-3488 [Abstract] [Full Text]  
  • Wunderlich, Z., Mirny, L. A. (2008). Spatial effects on the speed and reliability of protein-DNA search. Nucleic Acids Res 36: 3570-3578 [Abstract] [Full Text]  
  • Schulmeister, S., Ruttorf, M., Thiem, S., Kentner, D., Lebiedz, D., Sourjik, V. (2008). Protein exchange dynamics at chemoreceptor clusters in Escherichia coli. Proc. Natl. Acad. Sci. USA 105: 6403-6408 [Abstract] [Full Text]  
  • White, A. P., Gibson, D. L., Grassl, G. A., Kay, W. W., Finlay, B. B., Vallance, B. A., Surette, M. G. (2008). Aggregation via the Red, Dry, and Rough Morphotype Is Not a Virulence Adaptation in Salmonella enterica Serovar Typhimurium. Infect. Immun. 76: 1048-1058 [Abstract] [Full Text]  
  • Lindner, A. B., Madden, R., Demarez, A., Stewart, E. J., Taddei, F. (2008). Asymmetric segregation of protein aggregates is associated with cellular aging and rejuvenation. Proc. Natl. Acad. Sci. USA 105: 3076-3081 [Abstract] [Full Text]  
  • Campbell, C. S., Mullins, R. D. (2007). In vivo visualization of type II plasmid segregation: bacterial actin filaments pushing plasmids. JCB 179: 1059-1066 [Abstract] [Full Text]  
  • Beg, Q. K., Vazquez, A., Ernst, J., de Menezes, M. A., Bar-Joseph, Z., Barabasi, A.-L., Oltvai, Z. N. (2007). Intracellular crowding defines the mode and sequence of substrate uptake by Escherichia coli and constrains its metabolic activity. Proc. Natl. Acad. Sci. USA 104: 12663-12668 [Abstract] [Full Text]  
  • Suel, G. M., Kulkarni, R. P., Dworkin, J., Garcia-Ojalvo, J., Elowitz, M. B. (2007). Tunability and Noise Dependence in Differentiation Dynamics. Science 315: 1716-1719 [Abstract] [Full Text]  
  • Sanford, C., Yip, M. L.K., White, C., Parkinson, J. (2006). Cell++--simulating biochemical pathways. Bioinformatics 22: 2918-2925 [Abstract] [Full Text]  
  • Shih, Y.-L., Rothfield, L. (2006). The Bacterial Cytoskeleton. Microbiol. Mol. Biol. Rev. 70: 729-754 [Abstract] [Full Text]  
  • Konopka, M. C., Shkel, I. A., Cayley, S., Record, M. T., Weisshaar, J. C. (2006). Crowding and confinement effects on protein diffusion in vivo.. J. Bacteriol. 188: 6115-6123 [Abstract] [Full Text]  
  • Rodriguez, J. V., Kaandorp, J. A., Dobrzynski, M., Blom, J. G. (2006). Spatial stochastic modelling of the phosphoenolpyruvate-dependent phosphotransferase (PTS) pathway in Escherichia coli. Bioinformatics 22: 1895-1901 [Abstract] [Full Text]  
  • Kim, S. Y., Gitai, Z., Kinkhabwala, A., Shapiro, L., Moerner, W. E. (2006). Single molecules of the bacterial actin MreB undergo directed treadmilling motion in Caulobacter crescentus. Proc. Natl. Acad. Sci. USA 103: 10929-10934 [Abstract] [Full Text]  
  • Mullineaux, C. W., Nenninger, A., Ray, N., Robinson, C. (2006). Diffusion of green fluorescent protein in three cell environments in Escherichia coli.. J. Bacteriol. 188: 3442-3448 [Abstract] [Full Text]  
  • Nolan, S., Cowan, A. E., Koppel, D. E., Jin, H., Grote, E. (2006). FUS1 Regulates the Opening and Expansion of Fusion Pores between Mating Yeast. Mol. Biol. Cell 17: 2439-2450 [Abstract] [Full Text]  
  • Judd, E. M., Comolli, L. R., Chen, J. C., Downing, K. H., Moerner, W. E., McAdams, H. H. (2005). Distinct Constrictive Processes, Separated in Time and Space, Divide Caulobacter Inner and Outer Membranes. J. Bacteriol. 187: 6874-6882 [Abstract] [Full Text]  
  • Bialek, W., Setayeshgar, S. (2005). Physical limits to biochemical signaling. Proc. Natl. Acad. Sci. USA 102: 10040-10045 [Abstract] [Full Text]  
  • Doubrovinski, K., Howard, M. (2005). Stochastic model for Soj relocation dynamics in Bacillus subtilis. Proc. Natl. Acad. Sci. USA 102: 9808-9813 [Abstract] [Full Text]  
  • Ray, N., Nenninger, A., Mullineaux, C. W., Robinson, C. (2005). Location and Mobility of Twin Arginine Translocase Subunits in the Escherichia coli Plasma Membrane. J. Biol. Chem. 280: 17961-17968 [Abstract] [Full Text]  
  • Drew, D. A., Osborn, M. J., Rothfield, L. I. (2005). A polymerization-depolymerization model that accurately generates the self-sustained oscillatory system involved in bacterial division site placement. Proc. Natl. Acad. Sci. USA 102: 6114-6118 [Abstract] [Full Text]  
  • Lipkow, K., Andrews, S. S., Bray, D. (2005). Simulated Diffusion of Phosphorylated CheY through the Cytoplasm of Escherichia coli. J. Bacteriol. 187: 45-53 [Abstract] [Full Text]  
  • Zhou, B.-R., Liang, Y., Du, F., Zhou, Z., Chen, J. (2004). Mixed Macromolecular Crowding Accelerates the Oxidative Refolding of Reduced, Denatured Lysozyme: IMPLICATIONS FOR PROTEIN FOLDING IN INTRACELLULAR ENVIRONMENTS. J. Biol. Chem. 279: 55109-55116 [Abstract] [Full Text]  
  • Deich, J., Judd, E. M., McAdams, H. H., Moerner, W. E. (2004). Visualization of the movement of single histidine kinase molecules in live Caulobacter cells. Proc. Natl. Acad. Sci. USA 101: 15921-15926 [Abstract] [Full Text]  
  • Brehm-Stecher, B. F., Johnson, E. A. (2004). Single-Cell Microbiology: Tools, Technologies, and Applications. Microbiol. Mol. Biol. Rev. 68: 538-559 [Abstract] [Full Text]  
  • Anderson, D. E., Gueiros-Filho, F. J., Erickson, H. P. (2004). Assembly Dynamics of FtsZ Rings in Bacillus subtilis and Escherichia coli and Effects of FtsZ-Regulating Proteins. J. Bacteriol. 186: 5775-5781 [Abstract] [Full Text]  
  • Golding, I., Cox, E. C. (2004). RNA dynamics in live Escherichia coli cells. Proc. Natl. Acad. Sci. USA 101: 11310-11315 [Abstract] [Full Text]  
  • Mullineaux, C. W. (2004). FRAP analysis of photosynthetic membranes. J Exp Bot 55: 1207-1211 [Abstract] [Full Text]  
  • Judd, E. M., Ryan, K. R., Moerner, W. E., Shapiro, L., McAdams, H. H. (2003). Fluorescence bleaching reveals asymmetric compartment formation prior to cell division in Caulobacter. Proc. Natl. Acad. Sci. USA 100: 8235-8240 [Abstract] [Full Text]  
  • Cowan, A. E., Koppel, D. E., Setlow, B., Setlow, P. (2003). A soluble protein is immobile in dormant spores of Bacillus subtilis but is mobile in germinated spores: Implications for spore dormancy. Proc. Natl. Acad. Sci. USA 100: 4209-4214 [Abstract] [Full Text]  
  • Chakraborty, A., Das, I., Datta, R., Sen, B., Bhattacharyya, D., Mandal, C., Datta, A. K. (2002). A Single-domain Cyclophilin from Leishmania donovani Reactivates Soluble Aggregates of Adenosine Kinase by Isomerase-independent Chaperone Function. J. Biol. Chem. 277: 47451-47460 [Abstract] [Full Text]  
  • Dworkin, J., Losick, R. (2002). Does RNA polymerase help drive chromosome segregation in bacteria?. Proc. Natl. Acad. Sci. USA 99: 14089-14094 [Abstract] [Full Text]  
  • Gerland, U., Moroz, J. D., Hwa, T. (2002). Physical constraints and functional characteristics of transcription factor-DNA interaction. Proc. Natl. Acad. Sci. USA 99: 12015-12020 [Abstract] [Full Text]  
  • Meinhardt, H., de Boer, P. A. J. (2001). Pattern formation in Escherichia coli: A model for the pole-to-pole oscillations of Min proteins and the localization of the division site. Proc. Natl. Acad. Sci. USA 98: 14202-14207 [Abstract] [Full Text]  
  • Maki, N., Gestwicki, J. E., Lake, E. M., Kiessling, L. L., Adler, J. (2000). Motility and Chemotaxis of Filamentous Cells of Escherichia coli. J. Bacteriol. 182: 4337-4342 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»