Analysis of receptor-ligand interactions using nitrocellulose gel transfer: Application to Torpedo acetylcholine receptor and alpha-bungarotoxin
✍ Scribed by Barry Oblas; Norman D. Boyd; Robert H. Singer
- Publisher
- Elsevier Science
- Year
- 1983
- Tongue
- English
- Weight
- 748 KB
- Volume
- 130
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
A nitrocellulose-gel transfer technique has been adapted to study the interaction of a polypeptide ligand with individual receptor subunits. The acetylcholine receptor isolated from Torpedo californica has been separated into its subunits by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred in a renaturing environment to nitrocellulose sheets. The sheets were incubated with 'ZSI-alpha-bungarotoxin and autoradiographed. A single receptor polypeptide, the alpha subunit (40K) bound the labeled toxin. This binding was demonstrated to be both saturable and specific, although the affinity of '2SI-alpha-bungarotoxin (Kn, 165 nM) and the potency of d-tubocurarine to displace this binding (I&,, 1 mM) were both reduced by several orders of magnitude when compared to the native receptor.