Generation of coated intermediates of clathrin-mediated endocytosis on protein-free liposomes.
Clathrin-coated buds and dynamin-coated tubules morphologically similar to corresponding structures observed in synaptic membranes can be generated on protein-free liposomes by incubation with cytosol, or with clathrin coat proteins and purified dynamin, respectively. Dynamin- and clathrin-coated intermediates may form independently of each other, despite the coupling between the two processes typically observed in synaptic membranes. Formation of both structures on liposomes can occur in the absence of nucleotides. These findings indicate that interfaces between lipids and cytosolic proteins are fully sufficient to deform lipids bilayers into buds and tubules. They suggest that a main function of membrane proteins is to act as positive and negative regulators of coat assembly, therefore controlling these processes in time and space.
Takei
K
K
Department of Cell Biology and Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
Haucke
V
V
Slepnev
V
V
Farsad
K
K
Salazar
M
M
Chen
H
H
De Camilli
P
P
D000818:Animals; D001921:Brain; D002417:Cattle; D002451:Cell Compartmentation; D002474:Cell-Free System / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism*; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D034281:Dynamins; D004705:Endocytosis; D020558:GTP Phosphohydrolases / Q000302:isolation & purification; D006801:Humans; D008081:Liposomes / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000302:isolation & purification / Q000378:metabolism*; D009711:Nucleotides / Q000494:pharmacology; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification; D011485:Protein Binding; D051381:Rats; D013570:Synaptic Membranes / Q000378:metabolism
D002966:Clathrin; D008081:Liposomes; D009419:Nerve Tissue Proteins; D009711:Nucleotides; C075637:amphiphysin; C097702:synaptojanin; D010744:Phosphoric Monoester Hydrolases; D020558:GTP Phosphohydrolases; D034281:Dynamins