Constraining the Expression of Nicotinic Acetylcholine Receptors by Using Pentameric Constructs
Much of our understanding of ligand-gated ion channels comes from heterologous expression studies. However, this technique cannot produce receptors with a predetermined subunit composition for channels formed by several different subunits and cannot insert a single mutation copy if the subunit of in...
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Published in | Molecular pharmacology Vol. 69; no. 2; pp. 558 - 563 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
United States
American Society for Pharmacology and Experimental Therapeutics
01.02.2006
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Subjects | |
Online Access | Get full text |
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Summary: | Much of our understanding of ligand-gated ion channels comes from heterologous expression studies. However, this technique
cannot produce receptors with a predetermined subunit composition for channels formed by several different subunits and cannot
insert a single mutation copy if the subunit of interest is present in several copies in the channel. Here, we describe a
novel approach that overcomes these problems by expressing pentameric constructs, in which the code of the five subunits is
linked (i.e., β4_β4_α3_β4_α3). This is the first time that a concatemer of the complete pentameric receptor has been expressed
for channels in the cysteine-loop superfamily. The presence of the linker did not change the agonist or antagonist sensitivity
of α3β4 nicotinic receptors. We show evidence that the expressed receptors were made up of α3 and β4 subunits in one pentameric
fusion protein as designed in the construct. This approach can be applied to any nicotinic superfamily receptor to produce
channels with a defined subunit arrangement and to introduce specific mutations at any desired location of the pentameric
fusion protein. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0026-895X 1521-0111 |
DOI: | 10.1124/mol.105.019356 |