Influence of the clathrin coat on the membrane lipidic organization of endocytic vesicles: a fluorescence study.
Endocytic coated vesicles (CV) were purified from bovine brain, and uncoated vesicles (UV) were obtained from the latter by dialysis against 1 M Tris. Membrane dynamics were explored in both vesicle populations using two complementary fluorescence approaches: diphenylhexatriene fluorescence anisotropy to account for rotational lipid movements, and pyrene excimerization with a phosphatidylcholine pyrene derivative for translational motion. It was concluded that membrane fluidity was considerably higher in UV than in CV, and that adding bulk coat proteins (adaptors+clathrin) to uncoated vesicles re-established the low fluidity found in coated vesicles. However, adding coat protein constituents separately had no effect.
Liaubet
A
A
Centre de Biophysique Moléculaire, UPR 4301 du CNRS, Orléans, France.
Egret-Charlier
M
M
Kuhry
J G
JG
D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism*; D002966:Clathrin / Q000378:metabolism*; D004161:Diphenylhexatriene; D011992:Endosomes / Q000737:chemistry / Q000378:metabolism*; D005454:Fluorescence Polarization; D008560:Membrane Fluidity; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism*; D013696:Temperature
D002966:Clathrin; D008563:Membrane Lipids; D004161:Diphenylhexatriene