Freestanding lipid bilayers as substrates for electron cryomicroscopy of integral membrane proteins.
Phospholipid bilayers, 40 A thick, were generated as electron microscope substrates by submerging copper grids overlaid with holey plastic through a lipid monolayer on a water surface. Previously formed proteoliposomes containing single-particle membrane proteins in their bilayers were then fused into the newly formed bilayer substrate. To demonstrate this methodology, multi-drug resistance protein P-glycoprotein was incorporated into these bilayers and imaged by fixed beam microscopy and scanning transmission electron microscopy.
McAlduff
D E
DE
Ontario Cancer Institute and Department of Medical Biophysics, 610 University Avenue, University of Toronto, Toronto, Ontario, M5G 2M9, Canada.
Heng
Y M
YM
Ottensmeyer
F P
FP
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D020285:Cryoelectron Microscopy; D008051:Lipid Bilayers / Q000378:metabolism*; D008081:Liposomes; D008565:Membrane Proteins / Q000378:metabolism* / Q000648:ultrastructure*; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D010743:Phospholipids / Q000378:metabolism
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D008051:Lipid Bilayers; D008081:Liposomes; D008565:Membrane Proteins; D010743:Phospholipids