Probing solution structure of the pentameric ligand-gated ion channel GLIC by small-angle neutron scattering
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Probing solution structure of the pentameric ligand-gated ion channel GLIC by small-angle neutron scattering. / Lycksell, Marie; Rovsnik, Urska; Bergh, Cathrine; Johansen, Nicolai T.; Martel, Anne; Porcar, Lionel; Arleth, Lise; Howard, Rebecca J.; Lindahl, Erik.
I: Proceedings of the National Academy of Sciences of the United States of America, Bind 118, Nr. 37, 2108006118, 14.09.2021.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Probing solution structure of the pentameric ligand-gated ion channel GLIC by small-angle neutron scattering
AU - Lycksell, Marie
AU - Rovsnik, Urska
AU - Bergh, Cathrine
AU - Johansen, Nicolai T.
AU - Martel, Anne
AU - Porcar, Lionel
AU - Arleth, Lise
AU - Howard, Rebecca J.
AU - Lindahl, Erik
PY - 2021/9/14
Y1 - 2021/9/14
N2 - Pentameric ligand-gated ion channels undergo subtle conformational cycling to control electrochemical signal transduction in many kingdoms of life. Several crystal structures have now been reported in this family, but the functional relevance of such models remains unclear. Here, we used small-angle neutron scattering (SANS) to probe ambient solution-phase properties of the pHgated bacterial ion channel GLIC under resting and activating conditions. Data collection was optimized by inline paused-flow size-exclusion chromatography, and exchanging into deuterated detergent to hide the micelle contribution. Resting-state GLIC was the best-fit crystal structure to SANS curves, with no evidence for divergent mechanisms. Moreover, enhanced-sampling moleculardynamics simulations enabled differential modeling in resting versus activating conditions, with the latter corresponding to an intermediate ensemble of both the extracellular and transmembrane domains. This work demonstrates state-dependent changes in a pentameric ion channel by SANS, an increasingly accessible method for macromolecular characterization with the coming generation of neutron sources.
AB - Pentameric ligand-gated ion channels undergo subtle conformational cycling to control electrochemical signal transduction in many kingdoms of life. Several crystal structures have now been reported in this family, but the functional relevance of such models remains unclear. Here, we used small-angle neutron scattering (SANS) to probe ambient solution-phase properties of the pHgated bacterial ion channel GLIC under resting and activating conditions. Data collection was optimized by inline paused-flow size-exclusion chromatography, and exchanging into deuterated detergent to hide the micelle contribution. Resting-state GLIC was the best-fit crystal structure to SANS curves, with no evidence for divergent mechanisms. Moreover, enhanced-sampling moleculardynamics simulations enabled differential modeling in resting versus activating conditions, with the latter corresponding to an intermediate ensemble of both the extracellular and transmembrane domains. This work demonstrates state-dependent changes in a pentameric ion channel by SANS, an increasingly accessible method for macromolecular characterization with the coming generation of neutron sources.
KW - Cys-loop receptors
KW - gating
KW - small-angle neutron scattering
KW - molecular dynamics
KW - deuterated detergent
KW - X-RAY-STRUCTURE
KW - ACETYLCHOLINE-RECEPTOR
KW - GLOEOBACTER-VIOLACEUS
KW - MEMBRANE-PROTEINS
KW - OPEN PROBABILITY
KW - OPEN STATE
KW - SEC-SANS
KW - MECHANISM
KW - PORE
KW - CONFORMATION
U2 - 10.1073/pnas.2108006118
DO - 10.1073/pnas.2108006118
M3 - Journal article
C2 - 34504004
VL - 118
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 37
M1 - 2108006118
ER -
ID: 283457345