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Journal of Bacteriology, May 2006, p. 3442-3448, Vol. 188, No. 10
0021-9193/06/$08.00+0     doi:10.1128/JB.188.10.3442-3448.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Diffusion of Green Fluorescent Protein in Three Cell Environments in Escherichia Coli

Conrad W. Mullineaux,1* Anja Nenninger,2 Nicola Ray,2 and Colin Robinson2

School of Biological and Chemical Sciences, Queen Mary, University of London, Mile End Road, London E1 4NS, United Kingdom,1 Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, United Kingdom2

Received 25 November 2005/ Accepted 1 March 2006

Surprisingly little is known about the physical environment inside a prokaryotic cell. Knowledge of the rates at which proteins and other cell components can diffuse is crucial for the understanding of a cell as a physical system. There have been numerous measurements of diffusion coefficients in eukaryotic cells by using fluorescence recovery after photobleaching (FRAP) and related techniques. Much less information is available about diffusion coefficients in prokaryotic cells, which differ from eukaryotic cells in a number of significant respects. We have used FRAP to observe the diffusion of green fluorescent protein (GFP) in cells of Escherichia coli elongated by growth in the presence of cephalexin. GFP was expressed in the cytoplasm, exported into the periplasm using the twin-arginine translocation (Tat) system, or fused to an integral plasma membrane protein (TatA). We show that TatA-GFP diffuses in the plasma membrane with a diffusion coefficient comparable to that of a typical eukaryotic membrane protein. A previous report showed a very low rate of protein diffusion in the E. coli periplasm. However, we measured a GFP diffusion coefficient only slightly smaller in the periplasm than that in the cytoplasm, showing that both cell compartments are relatively fluid environments.

* Corresponding author. Mailing address: School of Biological and Chemical Sciences, Queen Mary, University of London, Mile End Road, London E1 4NS, United Kingdom. Phone: 44 20 7882 7008. Fax: 44 20 8983 0973. E-mail: c.mullineaux{at}qmul.ac.uk.

Journal of Bacteriology, May 2006, p. 3442-3448, Vol. 188, No. 10
0021-9193/06/$08.00+0     doi:10.1128/JB.188.10.3442-3448.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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