Anomalous protein diffusion in living cells as seen by fluorescence correlation spectroscopy.
We investigate the challenges and limitations that are encountered when studying membrane protein dynamics in vivo by means of fluorescence correlation spectroscopy (FCS). Based on theoretical arguments and computer simulations, we show that, in general, the fluctuating fluorescence has a fractal dimension D(0) >or= 1.5, which is determined by the anomality alpha of the diffusional motion of the labeled particles, i.e., by the growth of their mean square displacement as (Deltax)(2) approximately t(alpha). The fractality enforces an initial power-law behavior of the autocorrelation function and related quantities for small times. Using this information, we show by FCS that Golgi resident membrane proteins move subdiffusively in the endoplasmic reticulum and the Golgi apparatus in vivo. Based on Monte Carlo simulations for FCS on curved surfaces, we can rule out that the observed anomalous diffusion is a result of the complex topology of the membrane. The apparent mobility of particles as determined by FCS, however, is shown to depend crucially on the shape of the membrane and its motion in time. Due to this fact, the hydrodynamic radius of the tracked particles can be easily overestimated by an order of magnitude.
Weiss
Matthias
M
Cell Biology and Cell Biophysics Programme, European Molecular Biology Laboratory, 69117 Heidelberg, Germany. mweiss@embl-heidelberg.de
Hashimoto
Hitoshi
H
Nilsson
Tommy
T
D000465:Algorithms; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D004058:Diffusion; D004721:Endoplasmic Reticulum / Q000378:metabolism* / Q000648:ultrastructure; D017709:Fractals; D006056:Golgi Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D008954:Models, Biological; D009038:Motion; D017350:N-Acetylgalactosaminyltransferases / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013223:Statistics as Topic; D014018:Tissue Distribution
D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D017350:N-Acetylgalactosaminyltransferases