Live-cell imaging of single receptor composition using zero-mode waveguide nanostructures.
We exploit the optical and spatial features of subwavelength nanostructures to examine individual receptors on the plasma membrane of living cells. Receptors were sequestered in portions of the membrane projected into zero-mode waveguides. Using single-step photobleaching of green fluorescent protein incorporated into individual subunits, the resulting spatial isolation was used to measure subunit stoichiometry in α4β4 and α4β2 nicotinic acetylcholine and P2X2 ATP receptors. We also show that nicotine and cytisine have differential effects on α4β2 stoichiometry.
Division of Biology 156-29, California Institute of Technology, 1200 East California Boulevard, Pasadena, California 91125, United States.
Christopher I
CI
Richards
Khai
K
Luong
Rahul
R
Srinivasan
Stephen W
SW
Turner
Dennis A
DA
Dougherty
Jonas
J
Korlach
Henry A
HA
Lester
D000470:Alkaloids / Q000737:chemistry; D000818:Animals; D001392:Azocines / Q000737:chemistry; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000737:chemistry*; D051379:Mice; D049329:Nanostructures / Q000737:chemistry*; D009538:Nicotine / Q000737:chemistry; D010316:Particle Size; D011807:Quinolizines / Q000737:chemistry; D011978:Receptors, Nicotinic / Q000737:chemistry*; D058476:Receptors, Purinergic P2X2 / Q000737:chemistry*; D013499:Surface Properties; D000093843:Quinolizidine Alkaloids
D000470:Alkaloids; D001392:Azocines; D049452:Green Fluorescent Proteins; D009538:Nicotine; D011807:Quinolizines; D011978:Receptors, Nicotinic; D058476:Receptors, Purinergic P2X2; C004712:cytisine; D000093843:Quinolizidine Alkaloids