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Science 21 April 1995:
Vol. 268. no. 5209, pp. 439 - 442
DOI: 10.1126/science.7716551
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Articles

Science, Vol 268, Issue 5209, 439-442
Copyright © 1995 by American Association for the Advancement of Science

articles

Nicotinic receptor binding site probed with unnatural amino acid incorporation in intact cells

MW Nowak, PC Kearney, Sampson JR, ME Saks, CG Labarca, SK Silverman, W Zhong, J Thorson, JN Abelson, N Davidson, and al. et

Division of Biology, California Institute of Technology, Pasadena 91125, USA.

The nonsense codon suppression method for unnatural amino acid incorporation has been applied to intact cells and combined with electrophysiological analysis to probe structure-function relations in the nicotinic acetylcholine receptor. Functional receptors were expressed in Xenopus oocytes when tyrosine and phenylalanine derivatives were incorporated at positions 93, 190, and 198 in the binding site of the alpha subunit. Subtle changes in the structure of an individual side chain produced readily detectable changes in the function of this large channel protein. At each position, distinct features of side chain structure dominated the dose-response relation, probably by governing the agonist-receptor binding.


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