A substituted cysteine accessibility protocol to probe orthosteric and non-canonical ligand binding to ion channels.

Authors

Gauri C Kulkarni, Christian J Peters

Published in

STAR protocols. Volume 7. Issue 4. Pages 104934. Nov 02, 2026. Epub Nov 02, 2026.

Abstract

The Substituted Cysteine Accessibility Method (SCAM) is an approach for probing the biophysics and pharmacology of ion channels by targeting covalent modifications to individual amino acids. Here, we present a protocol to constrain channel stoichiometry and identify asymmetric or non-canonical ligand-binding sites using SCAM followed by electrophysiology in mammalian heterologous expression systems, in the context of heteropentameric nicotinic acetylcholine receptors. This protocol is applicable to examination of multiple channel properties, including ligand binding, gating, pore conductance, and selectivity. For complete information on the use and execution of this protocol, please refer to Kulkarni et al.¹.

PMID:
42970437
Bibliographic data and abstract were imported from PubMed on 03 Nov 2026.

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