Amino acid determinants of alpha 7 nicotinic acetylcholine receptor surface expression.
Transient transfection has not been a successful method to express the alpha7 nicotinic acetylcholine receptor such that these receptors are detected on the cell surface. This is not the case for all ligand-gated ion channels. Transient transfection with the 5-hydroxytryptamine type 3 subunit cDNA results in detectable surface receptor expression. Cell lines stably expressing the alpha7 nicotinic acetylcholine receptor produce detectable, albeit variable, levels of surface receptor expression. alpha7 nicotinic acetylcholine receptor surface expression is dependent, at least in part, on cell-specific factors. In addition to factors provided by the cells used for receptor expression, we hypothesize that the surface expression level in transfected cells is an intrinsic property of the receptor protein under study. Employing a set of alpha7-5-hydroxytryptamine type 3 chimeric receptor subunit cDNAs, we expressed these constructs in a transient transfection system and quantified surface receptor expression. We have identified amino acids that control receptor distribution between surface and intracellular pools; surface receptor expression can be manipulated without affecting the total number of receptors. These determinants function independently of the cell line used for expression and the transfection method employed. How these surface expression determinants in the alpha7 nicotinic acetylcholine receptor might influence synaptic efficacy is discussed.
Dineley
K T
KT
Division of Neuroscience, Baylor College of Medicine, Houston, Texas 77030, USA. kdineley@sensor.neusc.bcm.tmc.edu
Patrick
J W
JW
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000596:Amino Acids; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D008969:Molecular Sequence Data; D011978:Receptors, Nicotinic / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D000596:Amino Acids; D018076:DNA, Complementary; D011978:Receptors, Nicotinic