Diffusion coefficients of GFP in cell

Range amoeba Dictyostelium discoideum 24: Swiss mouse NIH-3T3 fibroblasts 27 µm^2/sec
Organism Eukaryotes
Reference Mika JT, Poolman B. Macromolecule diffusion and confinement in prokaryotic cells. Curr Opin Biotechnol. 2011 Feb22(1):117-26. doi: 10.1016/j.copbio.2010.09.009 p.118 right columnPubMed ID20952181
Primary Source [12] Potma EO et al., Reduced protein diffusion rate by cytoskeleton in vegetative and polarized dictyostelium cells. Biophys J. 2001 Oct81(4):2010-9 [13] Swaminathan R, Hoang CP, Verkman AS. Photobleaching recovery and anisotropy decay of green fluorescent protein GFP-S65T in solution and cells: cytoplasmic viscosity probed by green fluorescent protein translational and rotational diffusion. Biophys J. 1997 Apr72(4):1900-7. DOI: 10.1016/S0006-3495(97)78835-0PubMed ID11566774, 9083693
Method Abstract: "[Investigators] review recent observations on the mobility of macromolecules and their spatial organization in live bacterial cells. [They] outline the major fluorescence microscopy-based methods to determine the mobility and thus the diffusion coefficients (D) of molecules, which is not trivial in small cells." Primary source [12] abstract: "Fluorescence recovery after photobleaching measurements with high spatial resolution are performed to elucidate the impact of the actin cytoskeleton on translational mobility of green fluorescent protein (GFP) in aqueous domains of Dictyostelium discoideum amoebae." Primary source [13] abstract: "The green fluorescent protein (GFP) was used as a noninvasive probe to quantify the rheological properties of cell cytoplasm. GFP mutant S65T was purified from recombinant bacteria for solution studies, and expressed in CHO cell cytoplasm."
Comments P.118 right column: "The mobility of GFP in live E. coli cells is an order of magnitude slower than that in diluted aqueous solutions (87 μm^2/s [primary source 12]) and also slower than that in eukaryotic cytoplasm (e.g. 27 μm^2/s in Swiss 3T3 fibroblasts [primary source 13] or 24 μm^2/s [primary source 12] in D. discoideum). The faster diffusion coefficient in D. discoideum may reflect the much lower RNA concentration as the protein crowding seems similar to that of prokaryotic cells, whereas in higher eukaryotes both the protein and nucleic acid concentrations are lower than in bacteria."
Entered by Uri M
ID 115863