Disparate modulation of plasma membrane protein lateral mobility by various cell permeabilizing agents.
The mobility of a cell surface protein on cells osmotically swollen by treatment with several different cell permeabilizing agents retains specific restraints despite detachment of the plasma membrane from the cortical cytoskeleton. Fluorescence photobleaching recovery experiments indicate that the lateral diffusion constants of immunoglobulin E (IgE)-receptor complexes on the surface of rat basophilic leukemia cells increase 2-5x following permeabilization with streptolysin O or digitonin, with little change in their mobile fractions. Swelling by hypo-osmotic treatment in water enhances lateral diffusion of IgE-receptor complexes and raises the mobile fractions to near 100%. In contrast, swelling by treatment with filipin arrests lateral diffusion, although rotational mobility remains unhindered. Lateral mobility of a fluorescent lipid analogue remains unchanged under these conditions. Crosslinking by anti-IgE antibodies redistributes the IgE-receptor complexes into large patches on untreated cells and on cells swollen by permeabilization with streptolysin O or digitonin, but not on cells swollen by treatment with filipin. The results indicate a diversity of effects of the various permeabilizing agents on the mobility of membrane proteins. In particular, treatment with filipin appears to reorganize the plasma membrane into a network of fluid domains on a scale smaller than the bleaching spot size used (approximately 1.5 microns).
Feder
T J
TJ
Department of Physics, Cornell University, Ithaca, New York 14853.
Chang
E Y
EY
Holowka
D
D
Webb
W W
WW
D000818:Animals; D001426:Bacterial Proteins; D001692:Biological Transport / Q000187:drug effects; D002463:Cell Membrane Permeability; D004058:Diffusion; D004072:Digitonin / Q000494:pharmacology*; D005372:Filipin / Q000494:pharmacology*; D019793:Fluorescein; D005452:Fluoresceins; D008565:Membrane Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence / Q000379:methods; D009995:Osmosis; D051381:Rats; D017455:Receptors, IgE / Q000378:metabolism; D013301:Streptolysins / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D014867:Water / Q000378:metabolism
D001426:Bacterial Proteins; D005452:Fluoresceins; D008565:Membrane Proteins; D017455:Receptors, IgE; D013301:Streptolysins; C025964:streptolysin O; D014867:Water; D005372:Filipin; D004072:Digitonin; D019793:Fluorescein