Clathrin-dependent and clathrin-independent endocytosis are differentially sensitive to insertion of poly (ethylene glycol)-derivatized cholesterol in the plasma membrane.
We examined the effect of a cholesterol derivative, poly (ethylene glycol) cholesteryl ether on the structure/function of clathrin-coated pits and caveolae. Addition of the compound to cultured cells induced progressive smoothening of the surface. Markedly, when the incorporated amount exceeded 10% equivalent of the surface area, fluid pinocytosis, but not endocytosis of transferrin, became inhibited in K562 cells. In A431 cells, both clathrin-independent fluid phase uptake and the internalization of fluorescent cholera-toxin B through caveolae were inhibited with concomitant flattening of caveolae. In contrast, clathrin-mediated internalization of transferrin was not affected until the incorporated poly (ethylene glycol) cholesteryl ether exceeded 20% equivalent of the plasma membrane surface area, at which point opened clathrin-coated pits accumulated. The cells were ruptured upon further addition of poly (ethylene glycol) cholesteryl ether. We propose that the primary reason for the differential effect of poly (ethylene glycol) cholesteryl ether is that the bulk membrane phase and caveolae are both more elastic than the rigid clathrin-coated pits. We analyzed the results with the current mechanical model (Rauch and Farge, Biophys J 2000;78:3036-3047) and suggest here that the functional clathrin-lattice is much stiffer than typical phospholipid bilayers.
Baba
T
T
Department of Anatomy, Yamanashi Medical University, Yamanashi 409-3898, Japan.
Rauch
C
C
Xue
M
M
Terada
N
N
Fujii
Y
Y
Ueda
H
H
Takayama
I
I
Ohno
S
S
Farge
E
E
Sato
S B
SB
D001692:Biological Transport / Q000187:drug effects; D021941:Caveolae / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D002772:Cholera Toxin / Q000493:pharmacokinetics*; D002784:Cholesterol / Q000031:analogs & derivatives / Q000097:blood / Q000737:chemistry / Q000494:pharmacology*; D002966:Clathrin / Q000378:metabolism*; D003034:Coated Pits, Cell-Membrane / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure*; D005613:Freeze Etching; D006801:Humans; D020014:K562 Cells; D007700:Kinetics; D010873:Pinocytosis / Q000187:drug effects / Q000502:physiology; D011092:Polyethylene Glycols / Q000737:chemistry / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D002966:Clathrin; C118255:polyethylene glycol-cholesterol; D011092:Polyethylene Glycols; D002772:Cholera Toxin; D002784:Cholesterol