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Biophys J, February 2000, p. 901-907, Vol. 78, No. 2

Translational Diffusion of Globular Proteins in the Cytoplasm of Cultured Muscle Cells

Martine Arrio-Dupont,* Georges Foucault,* Monique Vacher,* Philippe F. Devaux,dagger and Sophie Cribierdagger

 *Gènes et Protéines Musculaires, EP CNRS 1088, F91405 Orsay, and  dagger Institut de Biologie Physico-Chimique, UPR CNRS 9052, F75005 Paris, France

Modulated fringe pattern photobleaching (MFPP) was used to measure the translational diffusion of microinjected fluorescein isothiocyanate (FITC)-labeled proteins of different sizes in the cytoplasm of cultured muscle cells. This technique, which is an extension of the classical fluorescence recovery after photobleaching (FRAP) technique, allows the measurement of the translational diffusion of macromolecules over several microns. Proteins used had molecular masses between 21 and 540 kDa. The results clearly indicated that the diffusivity of the various proteins is a decreasing function of their hydrodynamic radius. This decrease is more rapid with globular proteins than with FITC-labeled dextrans (Arrio-Dupont et al., 1996, Biophys. J. 70:2327-2332), most likely because, unlike globular proteins, dextrans are randomly coiled macromolecules with a flexible structure. These data do not exclude the possibility of a rapid diffusion over a short distance, unobservable with our experimental set-up, which would take place within the first milliseconds after bleaching and would correspond to the diffusion in restricted domains followed by impeded diffusion provoked by the network of microtubules, microfilaments, and intermediate filaments. Thus our results may complement rather than contradict those of Verkman and collaborators (Seksek et al., 1997, J. Cell Biol. 138:1-12). The biological consequence of the size-dependent restriction of the mobility of proteins in the cell cytoplasm is that the formation of intracellular complexes with other proteins considerably reduces their mobility.

Biophys J, February 2000, p. 901-907, Vol. 78, No. 2
© 2000 by the Biophysical Society   0006-3495/00/02/901/07  $2.00


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