Membrane lipid response to clathrin coat protein determined by infrared spectroscopy. Possible involvement in coated vesicle formation.
Clathrin, the major structural protein associated with both coated pits and coated vesicles, has been implicated in the dynamics of various endocytotic processes. In an attempt to define the mechanisms involved in the transition from uncoated membranes to clathrin-coated pits and then to coated vesicles, we investigated by infrared spectroscopy the lipid perturbations arising from the interactions of the clathrin coat with the bilayers of intact membrane assemblies. A comparison of the lipid acyl chain symmetric methylene stretching modes at approximately 2850 cm-1 for isolated clathrin-coated vesicles, uncoated vesicles, and synaptic membranes at 21, 38, and 50 degrees C indicated that clathrin significantly increases the number of gauche chain conformers in the bilayer matrix of the coated vesicle system. The increase in lipid disorder at 21 degrees C, accompanying the observed 0.44-cm-1 frequency increase for coated vesicles compared to uncoated vesicles, is approximately equivalent to the intrachain disorder incurred in heating liquid crystalline dimyristoylphosphatidylcholine multilayers by approximately 10 degrees C. The implications of these results on coated vesicle formation are discussed.
D000818:Animals; D001921:Brain / Q000378:metabolism; D002417:Cattle; D002458:Cell Fractionation; D002966:Clathrin / Q000378:metabolism*; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism* / Q000648:ultrastructure; D011992:Endosomes / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism*; D011485:Protein Binding; D013055:Spectrophotometry, Infrared; D013570:Synaptic Membranes / Q000378:metabolism* / Q000648:ultrastructure
D002966:Clathrin; D008563:Membrane Lipids