Compartmentalized structure of the plasma membrane for receptor movements as revealed by a nanometer-level motion analysis.
Movements of transferrin and alpha 2-macroglobulin receptor molecules in the plasma membrane of cultured normal rat kidney (NRK) fibroblastic cells were investigated by video-enhanced contrast optical microscopy with 1.8 nm spatial precision and 33 ms temporal resolution by labeling the receptors with the ligand-coated nanometer-sized colloidal gold particles. For both receptor species, most of the movement trajectories are of the confined diffusion type, within domains of approximately 0.25 microns2 (500-700 nm in diagonal length). Movement within the domains is random with a diffusion coefficient approximately 10(-9) cm2/s, which is consistent with that expected for free Brownian diffusion of proteins in the plasma membrane. The receptor molecules move from one domain to one of the adjacent domains at an average frequency of 0.034 s-1 (the residence time within a domain approximately 29 s), indicating that the plasma membrane is compartmentalized for diffusion of membrane receptors and that long-range diffusion is the result of successive intercompartmental jumps. The macroscopic diffusion coefficients for these two receptor molecules calculated on the basis of the compartment size and the intercompartmental jump rate are approximately 2.4 x 10(-11) cm2/s, which is consistent with those determined by averaging the long-term movements of many particles. Partial destruction of the cytoskeleton decreased the confined diffusion mode, increased the simple diffusion mode, and induced the directed diffusion (transport) mode. These results suggest that the boundaries between compartments are made of dynamically fluctuating membrane skeletons (membrane-skeleton fence model).
Sako
Y
Y
Department of Life Sciences, University of Tokyo, Japan.
Kusumi
A
A
D000818:Animals; D002451:Cell Compartmentation / Q000187:drug effects; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D002478:Cells, Cultured; D015638:Cytochalasin D / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects; D004058:Diffusion; D005347:Fibroblasts; D017979:Gold Colloid; D006651:Histocytochemistry; D007668:Kidney / Q000166:cytology; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008857:Microscopy, Interference; D008954:Models, Biological; D008956:Models, Chemical; D009038:Motion; D051381:Rats; D011971:Receptors, Immunologic / Q000378:metabolism*; D011990:Receptors, Transferrin / Q000378:metabolism*; D014741:Video Recording; D014747:Vinblastine / Q000494:pharmacology
D017979:Gold Colloid; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D011971:Receptors, Immunologic; D011990:Receptors, Transferrin; D015638:Cytochalasin D; D014747:Vinblastine