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@ARTICLE{Fahlke:885449,
author = {Fahlke, Christoph and Balandin, Taras and Astashkin, Roman
and Alleva, Claudia and Machtens, Jan-Philipp and Gordeliy,
Valentin and Baeken, Christian and Kovalev, Kirill and
Berndt, Meike},
title = {{N}a + -dependent gate dynamics and electrostatic
attraction ensure substrate coupling in glutamate
transporters},
journal = {Science advances},
volume = {6},
number = {47},
issn = {2375-2548},
address = {Washington, DC [u.a.]},
publisher = {Assoc.},
reportid = {FZJ-2020-03832},
pages = {eaba9854 -},
year = {2020},
abstract = {Excitatory amino acid transporters (EAATs) harness [Na+],
[K+], and [H+] gradients for fast and efficient glutamate
removal from the synaptic cleft. Since each glutamate is
cotransported with three Na+ ions, [Na+] gradients are the
predominant driving force for glutamate uptake. We combined
all-atom molecular dynamics simulations, fluorescence
spectroscopy, and x-ray crystallography to study
Na+:substrate coupling in the EAAT homolog GltPh. A lipidic
cubic phase x-ray crystal structure of wild-type, Na+-only
bound GltPh at 2.5-Å resolution revealed the fully open,
outward-facing state primed for subsequent substrate
binding. Simulations and kinetic experiments established
that only the binding of two Na+ ions to the Na1 and Na3
sites ensures complete HP2 gate opening via a conformational
selection-like mechanism and enables high-affinity substrate
binding via electrostatic attraction. The combination of
Na+-stabilized gate opening and electrostatic coupling of
aspartate to Na+ binding provides a constant Na+:substrate
transport stoichiometry over a broad range of
neurotransmitter concentrations.},
cin = {IBI-1 / IBI-7 / JARA-HPC},
ddc = {500},
cid = {I:(DE-Juel1)IBI-1-20200312 / I:(DE-Juel1)IBI-7-20200312 /
$I:(DE-82)080012_20140620$},
pnm = {551 - Functional Macromolecules and Complexes (POF3-551) /
MOLECULAR MODELLING OF BIFUNCTIONAL MEMBRANE TRANSPORT
PROTEINS $(jics40_20190501)$ / Molecular dynamics
simulations of P2X receptors $(jara0180_20200501)$ /
Multiscale simulations of voltage-gated sodium channel
complexes and clusters $(jics42_20191101)$ / Molecular
dynamics of the SLC26 family of ion channels and
transporters $(jara0177_20191101)$},
pid = {G:(DE-HGF)POF3-551 / $G:(DE-Juel1)jics40_20190501$ /
$G:(DE-Juel1)jara0180_20200501$ /
$G:(DE-Juel1)jics42_20191101$ /
$G:(DE-Juel1)jara0177_20191101$},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:33208356},
UT = {WOS:000592173500003},
doi = {10.1126/sciadv.aba9854},
url = {https://juser.fz-juelich.de/record/885449},
}