Allosteric binding site in a Cys-loop receptor ligand-binding
domain unveiled in the crystal structure of ELIC in complex
with chlorpromazine

J 2016

Allosteric binding site in a Cys-loop receptor ligand-binding domain unveiled in the crystal structure of ELIC in complex with chlorpromazine

NYS, Mieke, Eveline WIJCKMANS, Ana FARINHA, Ozge YOLUK, Magnus ANDERSSON et. al.

Základní údaje

Originální název

Allosteric binding site in a Cys-loop receptor ligand-binding domain unveiled in the crystal structure of ELIC in complex with chlorpromazine

Autoři

NYS, Mieke (56 Belgie), Eveline WIJCKMANS (56 Belgie), Ana FARINHA (56 Belgie), Ozge YOLUK (752 Švédsko), Magnus ANDERSSON (752 Švédsko), Marijke BRAMS (56 Belgie), Radovan SPURNÝ (703 Slovensko, garant, domácí), Steve PEIGNEUR (56 Belgie), Jan TYTGAT (56 Belgie), Erik LINDAHL (752 Švédsko) a Chris ULENS (56 Belgie)

Vydání

Proceedings of the National Academy of Sciences of the United States of America, WASHINGTON, National Academy of Sciences, 2016, 0027-8424

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10403 Physical chemistry

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 9.661

Kód RIV

RIV/00216224:14740/16:00093868

Organizační jednotka

Středoevropský technologický institut

DOI

http://dx.doi.org/10.1073/pnas.1603101113

UT WoS

000386087100020

Klíčová slova anglicky

ligand-gated ion channel; X-ray crystallography; allosteric modulation; Cys-loop receptor; nicotinic acetylcholine receptor

Štítky

rivok

Změněno: 10. 3. 2017 10:11, Mgr. Eva Špillingová

Anotace

V originále

Pentameric ligand-gated ion channels or Cys-loop receptors are responsible for fast inhibitory or excitatory synaptic transmission. The antipsychotic compound chlorpromazine is a widely used tool to probe the ion channel pore of the nicotinic acetylcholine receptor, which is a prototypical Cys-loop receptor. In this study, we determine the molecular determinants of chlorpromazine binding in the Erwinia ligand-gated ion channel (ELIC). We report the X-ray crystal structures of ELIC in complex with chlorpromazine or its brominated derivative bromopromazine. Unexpectedly, we do not find a chlorpromazine molecule in the channel pore of ELIC, but behind the beta 8-beta 9 loop in the extracellular ligand-binding domain. The beta 8-beta 9 loop is localized downstream from the neurotransmitter binding site and plays an important role in coupling of ligand binding to channel opening. In combination with electrophysiological recordings from ELIC cysteine mutants and a thiol-reactive derivative of chlorpromazine, we demonstrate that chlorpromazine binding at the beta 8-beta 9 loop is responsible for receptor inhibition. We further use molecular-dynamics simulations to support the X-ray data and mutagenesis experiments. Together, these data unveil an allosteric binding site in the extracellular ligand-binding domain of ELIC. Our results extend on previous observations and further substantiate our understanding of a multisite model for allosteric modulation of Cys-loop receptors.

Citovat

NYS, Mieke, Eveline WIJCKMANS, Ana FARINHA, Ozge YOLUK, Magnus ANDERSSON, Marijke BRAMS, Radovan SPURNÝ, Steve PEIGNEUR, Jan TYTGAT, Erik LINDAHL a Chris ULENS. Allosteric binding site in a Cys-loop receptor ligand-binding domain unveiled in the crystal structure of ELIC in complex with chlorpromazine. Proceedings of the National Academy of Sciences of the United States of America. WASHINGTON: National Academy of Sciences, 2016, roč. 113, č. 43, s. "E6696"-"E6703", 8 s. ISSN 0027-8424. Dostupné z: https://dx.doi.org/10.1073/pnas.1603101113.

@article{1375144,
   author = {Nys, Mieke and Wijckmans, Eveline and Farinha, Ana and Yoluk, Ozge and Andersson, Magnus and Brams, Marijke and Spurný, Radovan and Peigneur, Steve and Tytgat, Jan and Lindahl, Erik and Ulens, Chris},
   article_location = {WASHINGTON},
   article_number = {43},
   doi = {http://dx.doi.org/10.1073/pnas.1603101113},
   keywords = {ligand-gated ion channel; X-ray crystallography; allosteric modulation; Cys-loop receptor; nicotinic acetylcholine receptor},
   language = {eng},
   issn = {0027-8424},
   journal = {Proceedings of the National Academy of Sciences of the United States of America},
   title = {Allosteric binding site in a Cys-loop receptor ligand-binding domain unveiled in the crystal structure of ELIC in complex with chlorpromazine},
   url = {http://www.pnas.org/content/113/43/E6696},
   volume = {113},
   year = {2016}
}

TY  - JOUR
ID  - 1375144
AU  - Nys, Mieke - Wijckmans, Eveline - Farinha, Ana - Yoluk, Ozge - Andersson, Magnus - Brams, Marijke - Spurný, Radovan - Peigneur, Steve - Tytgat, Jan - Lindahl, Erik - Ulens, Chris
PY  - 2016
TI  - Allosteric binding site in a Cys-loop receptor ligand-binding domain unveiled in the crystal structure of ELIC in complex with chlorpromazine
JF  - Proceedings of the National Academy of Sciences of the United States of America
VL  - 113
IS  - 43
SP  - "E6696"-"E6703"
EP  - "E6696"-"E6703"
PB  - National Academy of Sciences
SN  - 00278424
KW  - ligand-gated ion channel
KW  - X-ray crystallography
KW  - allosteric modulation
KW  - Cys-loop receptor
KW  - nicotinic acetylcholine receptor
UR  - http://www.pnas.org/content/113/43/E6696
L2  - http://www.pnas.org/content/113/43/E6696
N2  - Pentameric ligand-gated ion channels or Cys-loop receptors are responsible for fast inhibitory or excitatory synaptic transmission. The antipsychotic compound chlorpromazine is a widely used tool to probe the ion channel pore of the nicotinic acetylcholine receptor, which is a prototypical Cys-loop receptor. In this study, we determine the molecular determinants of chlorpromazine binding in the Erwinia ligand-gated ion channel (ELIC). We report the X-ray crystal structures of ELIC in complex with chlorpromazine or its brominated derivative bromopromazine. Unexpectedly, we do not find a chlorpromazine molecule in the channel pore of ELIC, but behind the beta 8-beta 9 loop in the extracellular ligand-binding domain. The beta 8-beta 9 loop is localized downstream from the neurotransmitter binding site and plays an important role in coupling of ligand binding to channel opening. In combination with electrophysiological recordings from ELIC cysteine mutants and a thiol-reactive derivative of chlorpromazine, we demonstrate that chlorpromazine binding at the beta 8-beta 9 loop is responsible for receptor inhibition. We further use molecular-dynamics simulations to support the X-ray data and mutagenesis experiments. Together, these data unveil an allosteric binding site in the extracellular ligand-binding domain of ELIC. Our results extend on previous observations and further substantiate our understanding of a multisite model for allosteric modulation of Cys-loop receptors.
ER  -

NYS, Mieke, Eveline WIJCKMANS, Ana FARINHA, Ozge YOLUK, Magnus ANDERSSON, Marijke BRAMS, Radovan SPURNÝ, Steve PEIGNEUR, Jan TYTGAT, Erik LINDAHL a Chris ULENS. Allosteric binding site in a Cys-loop receptor ligand-binding domain unveiled in the crystal structure of ELIC in complex with chlorpromazine. \textit{Proceedings of the National Academy of Sciences of the United States of America}. WASHINGTON: National Academy of Sciences, 2016, roč.~113, č.~43, s.~''E6696''-''E6703'', 8 s. ISSN~0027-8424. Dostupné z: https://dx.doi.org/10.1073/pnas.1603101113.

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