6F12

GLIC mutant E181A

Summary for 6F12

• Entry DOI: 10.2210/pdb6f12/pdb
• Descriptor: Proton-gated ion channel, CHLORIDE ION, SODIUM ION, ... (7 entities in total)
• Functional Keywords: membrane protein, transport protein
• Biological source: Gloeobacter violaceus (strain PCC 7421)
• Cellular location: Cell inner membrane ; Multi- pass membrane protein : Q7NDN8
• Total number of polymer chains: 5
• Total formula weight: 186384.35

Authors

Hu, H.D.,Delarue, M. (deposition date: 2017-11-21, release date: 2018-01-10, Last modification date: 2024-05-08)

Primary citation

Nemecz, A.,Hu, H.,Fourati, Z.,Van Renterghem, C.,Delarue, M.,Corringer, P.J.
Full mutational mapping of titratable residues helps to identify proton-sensors involved in the control of channel gating in the Gloeobacter violaceus pentameric ligand-gated ion channel.
PLoS Biol., 15:e2004470-e2004470, 2017

PubMed Abstract: The Gloeobacter violaceus ligand-gated ion channel (GLIC) has been extensively studied by X-ray crystallography and other biophysical techniques. This provided key insights into the general gating mechanism of pentameric ligand-gated ion channel (pLGIC) signal transduction. However, the GLIC is activated by lowering the pH and the location of its putative proton activation site(s) still remain(s) unknown. To this end, every Asp, Glu, and His residue was mutated individually or in combination and investigated by electrophysiology. In addition to the mutational analysis, key mutations were structurally resolved to address whether particular residues contribute to proton sensing, or alternatively to GLIC-gating, independently of the side chain protonation. The data show that multiple residues located below the orthosteric site, notably E26, D32, E35, and D122 in the lower part of the extracellular domain (ECD), along with E222, H235, E243, and H277 in the transmembrane domain (TMD), alter GLIC activation. D122 and H235 were found to also alter GLIC expression. E35 is identified as a key proton-sensing residue, whereby neutralization of its side chain carboxylate stabilizes the active state. Thus, proton activation occurs allosterically to the orthosteric site, at the level of multiple loci with a key contribution of the coupling interface between the ECD and TMD.

PubMed: 29281623
DOI: 10.1371/journal.pbio.2004470

Experimental method

X-RAY DIFFRACTION (3.2 Å)