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Binding Sites and the Mechanism of Action of Propofol and a Photoreactive Analogue in Prokaryotic Voltage-Gated Sodium Channels
Journal article   Peer reviewed

Binding Sites and the Mechanism of Action of Propofol and a Photoreactive Analogue in Prokaryotic Voltage-Gated Sodium Channels

Elaine Yang, Weiming Bu, Antonio Suma, Vincenzo Carnevale, Kimberly C. Grasty, Patrick J. Loll, Kellie Woll, Natarajan Bhanu, Benjamin A. Garcia, Roderic G. Eckenhoff, …
ACS chemical neuroscience, v 12(20), pp 3898-3914
20 Oct 2021
PMID: 34607428

Abstract

Biochemistry & Molecular Biology Chemistry, Medicinal Life Sciences & Biomedicine Neurosciences Neurosciences & Neurology Pharmacology & Pharmacy Science & Technology
Propofol, one of the most commonly used intravenous general anesthetics, modulates neuronal function by interacting with ion channels. The mechanisms that link propofol binding to the modulation of distinct ion channel states, however, are not understood. To tackle this problem, we investigated the prokaryotic ancestors of eukaryotic voltage-gated Na+ channels (Nays) using unbiased photoaffinity labeling (PAL) with a diazirine derivative of propofol (AziPm), electrophysiological methods, and mutagenesis. AziPm inhibits Nav function in a manner that is indistinguishable from that of the parent compound by promoting activation-coupled inactivation. In several replicates (8/9) involving NaChBac and NavMs, we found adducts at residues located at the C-terminal end of the S4 voltage sensor, the S4-S5 linker, and the N-terminal end of the S5 segment. However, the non-inactivating mutant NaChBac-T220A yielded adducts that were different from those found in the wild-type counterpart, which suggested state-dependent changes at the binding site. Then, using molecular dynamics simulations to further elucidate the structural basis of Nay modulation by propofol, we show that the S4 voltage sensors and the S4-S5 linkers shape two distinct propofol binding sites in a conformation-dependent manner. Supporting the PAL and MD simulation results, we also found that Ala mutations of a subset of adducted residues have distinct effects on gating modulation of NaChBac and NavMs by propofol. The results of this study provide direct insights into the structural basis of the mechanism through which propofol binding promotes activation-coupled inactivation to inhibit Nay channel function.

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Domestic collaboration
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Web of Science research areas
Biochemistry & Molecular Biology
Chemistry, Medicinal
Neurosciences
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