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J Biol Chem, Vol. 275, Issue 18, 13974-13985, May 5, 2000
From the Division of Neuroscience, Baylor College of Medicine,
Houston, Texas 77030
Transient transfection has not been a successful
method to express the
Amino Acid Determinants of
7 Nicotinic Acetylcholine Receptor
Surface Expression*
and
7 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 7
nicotinic acetylcholine receptor produce detectable, albeit variable,
levels of surface receptor expression. 7 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
7-5-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 7
nicotinic acetylcholine receptor might influence synaptic efficacy is discussed.
*
This work was supported by National Institutes of Health
Grants F31 DA05694 and NS 13546.The costs of publication of this article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Division of
Neuroscience, Rm. S603, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030. Tel.: 713-798-3086; Fax: 713-798-3946; E-mail: kdineley@sensor.neusc.bcm.tmc.edu.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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