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Biophys J. 2003 Jan;84(1):356-66.

Confined diffusion without fences of a g-protein-coupled receptor as revealed by single particle tracking.

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  • 1Institut de Pharmacologie et Biologie Structurale, CNRS UMR 5089, 205, route de Narbonne, 31077 Toulouse Cedex, France.

Abstract

Single particle tracking is a powerful tool for probing the organization and dynamics of the plasma membrane constituents. We used this technique to study the micro -opioid receptor belonging to the large family of the G-protein-coupled receptors involved with other partners in a signal transduction pathway. The specific labeling of the receptor coupled to a T7-tag at its N-terminus, stably expressed in fibroblastic cells, was achieved by colloidal gold coupled to a monoclonal anti T7-tag antibody. The lateral movements of the particles were followed by nanovideomicroscopy at 40 ms time resolution during 2 min with a spatial precision of 15 nm. The receptors were found to have either a slow or directed diffusion mode (10%) or a walking confined diffusion mode (90%) composed of a long-term random diffusion and a short-term confined diffusion, and corresponding to a diffusion confined within a domain that itself diffuses. The results indicate that the confinement is due to an effective harmonic potential generated by long-range attraction between the membrane proteins. A simple model for interacting membrane proteins diffusion is proposed that explains the variations with the domain size of the short-term and long-term diffusion coefficients.

PMID:
12524289
[PubMed - indexed for MEDLINE]
PMCID:
PMC1302617
Free PMC Article
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