Neuronal nicotinic acetylcholine receptors expressed in Xenopus oocytes have a pentameric quaternary structure.
We have determined the subunit stoichiometry of chicken neuronal nicotinic acetylcholine receptors expressed in Xenopus oocytes by quantitation of the amount of radioactivity in individual subunits of [35S] methionine-labeled receptors. The chicken neuronal nicotinic acetylcholine receptor appears to be a pentamer of two alpha 4 acetylcholine-binding subunits and three beta 2 structural subunits. We also show that these expressed receptors bind L-[3H]nicotine with high affinity, are transported to the surface of the oocyte outer membrane, and cosediment on sucrose gradients with acetylcholine receptors isolated from chicken brain. Using this unique and generally applicable method of determining subunit stoichiometry of receptors expressed in oocytes, we obtained the expected (alpha 1) 2 beta 1 gamma delta stoichiometry for muscle-type acetylcholine receptors assembled from coexpression of either Torpedo alpha 1 or human alpha 1 subunits, with Torpedo beta 1, gamma, and delta subunits.
Anand
R
R
Slak Institute for Biological Studies, San Diego, California 92186-5800.
Conroy
W G
WG
Schoepfer
R
R
Whiting
P
P
Lindstrom
J
J
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002645:Chickens; D006801:Humans; D046911:Macromolecular Substances; D008715:Methionine / Q000378:metabolism; D009132:Muscles / Q000378:metabolism*; D009474:Neurons / Q000502:physiology*; D009538:Nicotine / Q000378:metabolism; D009865:Oocytes / Q000502:physiology*; D011978:Receptors, Nicotinic / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D014101:Torpedo; D014981:Xenopus
D046911:Macromolecular Substances; D011978:Receptors, Nicotinic; D009538:Nicotine; D008715:Methionine