Molecular determinants underlying volume-regulated anion channel subunit-dependent oxidation sensitivity.
VRAC
oxidation
start methionine
volume regulation
Journal
The Journal of physiology
ISSN: 1469-7793
Titre abrégé: J Physiol
Pays: England
ID NLM: 0266262
Informations de publication
Date de publication:
09 2022
09 2022
Historique:
received:
12
05
2022
accepted:
15
07
2022
pubmed:
22
7
2022
medline:
9
9
2022
entrez:
21
7
2022
Statut:
ppublish
Résumé
The volume-regulated anion channel (VRAC) is formed by LRRC8 subunits. Besides their role in the maintenance of cell homeostasis, VRACs are critically involved in oxidative stress mechanisms: reactive oxygen species directly modulate VRACs in a subunit-dependent manner. It was reported that LRRC8A-LRRC8E heteromeric channels are activated by oxidation, whereas LRRC8A-LRRC8C heteromers are inhibited. Here we adopted chimeric- as well as concatemeric-based strategies to identify residues responsible for the divergent effect of oxidants. We identified two cysteines in the first two leucine rich repeats of LRRC8E, C424 and C448, as the targets of oxidation. Oxidation likely results in the formation of a disulfide bond between the two cysteines, which in turn induces a conformational change leading to channel activation. Additionally, we found that LRRC8C inhibition is caused by oxidation of the first methionine. We thus identified crucial molecular elements involved in channel activation, which are conceivably relevant in determining physiological ROS effects. KEY POINTS: Volume-regulated anion channels (VRACs) are heterohexameric complexes composed of an essential LRRC8A subunit and a variable number of LRRC8B-E subunits. VRACs are directly regulated by oxidation, with LRRC8A-LRRC8E heteromers being potentiated and LRRC8A-LRRC8C heteromers being inhibited by oxidation. We identified two LRRC8E specific intracellular cysteines that form a disulfide bond upon oxidation leading to LRRC8A-LRRC8E potentiation. Inhibition of LRRC8A-LRRC8C heteromers is mediated by the oxidation of the start methionine, being additionally dependent on the identity of the LRR domain. Besides providing physiological insights concerning the outcome of reactive oxygen species modulation, the results point to key structural elements involved in VRAC activation.
Identifiants
pubmed: 35861288
doi: 10.1113/JP283321
pmc: PMC9540897
doi:
Substances chimiques
Anions
0
Disulfides
0
Membrane Proteins
0
Reactive Oxygen Species
0
Methionine
AE28F7PNPL
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
3965-3982Informations de copyright
© 2022 The Authors. The Journal of Physiology published by John Wiley & Sons Ltd on behalf of The Physiological Society.
Références
J Gen Physiol. 2004 Sep;124(3):273-87
pubmed: 15337822
EMBO J. 2015 Dec 14;34(24):2993-3008
pubmed: 26530471
Biol Chem. 2015 Sep;396(9-10):975-90
pubmed: 25868000
Biotechniques. 1994 Nov;17(5):876-81
pubmed: 7840967
Science. 2014 May 9;344(6184):634-8
pubmed: 24790029
BMC Biochem. 2010 Jul 01;11:25
pubmed: 20594348
Elife. 2018 Aug 10;7:
pubmed: 30095067
Channels (Austin). 2017 May 4;11(3):254-260
pubmed: 28121479
J Biol Chem. 2018 Aug 31;293(35):13440-13451
pubmed: 29925591
J Biol Chem. 2013 Sep 13;288(37):26489-96
pubmed: 23861395
Biophys J. 2016 Oct 4;111(7):1429-1443
pubmed: 27705766
Biochemistry. 2010 Sep 7;49(35):7748-55
pubmed: 20712299
Int J Mol Sci. 2018 Mar 02;19(3):
pubmed: 29498698
Proc Natl Acad Sci U S A. 2004 Apr 27;101(17):6770-3
pubmed: 15096609
J Biol Chem. 2014 Jun 13;289(24):17124-31
pubmed: 24782309
Pharmacol Rev. 2019 Jan;71(1):49-88
pubmed: 30573636
Nature. 2018 Jun;558(7709):254-259
pubmed: 29769723
J Cell Sci. 2017 Mar 15;130(6):1122-1133
pubmed: 28193731
Immunity. 2020 May 19;52(5):767-781.e6
pubmed: 32277911
Cell Physiol Biochem. 2021 Feb 13;55(S1):41-56
pubmed: 33577730
Nat Rev Mol Cell Biol. 2016 May;17(5):293-307
pubmed: 27033257
J Physiol. 2017 Nov 1;595(21):6719-6733
pubmed: 28841766
Amino Acids. 2003 Dec;25(3-4):259-74
pubmed: 14661089
Commun Biol. 2020 May 15;3(1):240
pubmed: 32415200
Elife. 2019 Jun 18;8:
pubmed: 31210638
J Gen Physiol. 2019 Feb 4;151(2):100-117
pubmed: 30651298
J Gen Physiol. 2006 Jan;127(1):51-65
pubmed: 16380443
Elife. 2019 Feb 18;8:
pubmed: 30775971
Soc Gen Physiol Ser. 1988;43:281-301
pubmed: 2479106
Int J Mol Sci. 2021 Aug 04;22(16):
pubmed: 34445066
Cell. 2014 Apr 10;157(2):447-458
pubmed: 24725410
Cell Physiol Biochem. 2007;20(6):773-80
pubmed: 17982259
J Biol Chem. 2004 Apr 2;279(14):13301-4
pubmed: 14761962
J Neurosci. 2009 Jul 8;29(27):8621-9
pubmed: 19587267
Nat Struct Mol Biol. 2018 Sep;25(9):797-804
pubmed: 30127360
J Physiol. 1988 Aug;402:687-702
pubmed: 2466988
Cell Death Dis. 2019 Dec 5;10(12):925
pubmed: 31804464
Bioessays. 2012 Jul;34(7):551-60
pubmed: 22532330