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@ARTICLE{Fruergaard:486058,
author = {Fruergaard, Marlene U. and Dach, Ingrid and Andersen, Jacob
L. and Ozol, Mette and Shahsavar, Azadeh and Quistgaard,
Esben M. and Poulsen, Hanne and Fedosova, Natalya U. and
Nissen, Poul},
title = {{T}he {N}a$^+$,{K}$^+$-{ATP}ase in complex with beryllium
fluoride mimics an {ATP}ase phosphorylated state},
journal = {The journal of biological chemistry},
volume = {298},
number = {9},
issn = {0021-9258},
address = {Bethesda, MD.},
publisher = {American Soc. for Biochemistry and Molecular Biology},
reportid = {PUBDB-2022-07058},
pages = {102317},
year = {2022},
note = {ISSN 0021-9258 not unique: **2 hits**.},
abstract = {The Na$^+$,K$^+$-ATPase generates electrochemical gradients
of Na$^+$ and K$^+$ across the plasma membrane via a
functional cycle that includes various phosphoenzyme
intermediates. However, the structure and function of these
intermediates and how metal fluorides mimick them require
further investigation. Here, we describe a 4.0 Å resolution
crystal structure and functional properties of the pig
kidney Na$^+$,K$^+$-ATPase stabilized by the inhibitor
beryllium fluoride (denoted E2–BeF$_x$). E2–BeF$_x$ is
expected to mimic properties of the E2P phosphoenzyme, yet
with unknown characteristics of ion and ligand binding. The
structure resembles the E2P form obtained by phosphorylation
from inorganic phosphate (P$_i$) and stabilized by
cardiotonic steroids, including a low-affinity Mg$^{2+}$
site near ion binding site II. Our anomalous Fourier
analysis of the crystals soaked in Rb$^+$ (a K$^+$ congener)
followed by a low-resolution rigid-body refinement
(6.9–7.5 Å) revealed preocclusion transitions leading to
activation of the dephosphorylation reaction. We show that
the Mg$^{2+}$ location indicates a site of initial K$^+$
recognition and acceptance upon binding to the outward-open
E2P state after Na$^+$ release. Furthermore, using binding
and activity studies, we find that the BeF$_x$-inhibited
enzyme is also able to bind ADP/ATP and Na$^+$. These
results relate the E2–BeF$_x$ complex to a transient
K$^+$- and ADP-sensitive E∗P intermediate of the
functional cycle of the Na$^+$,K$^+$-ATPase, prior to E2P.},
cin = {EMBL-User},
ddc = {610},
cid = {I:(DE-H253)EMBL-User-20120814},
pnm = {6G3 - PETRA III (DESY) (POF4-6G3)},
pid = {G:(DE-HGF)POF4-6G3},
experiment = {EXP:(DE-H253)P-P14-20150101 / EXP:(DE-H253)P-P13-20150101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:35926706},
UT = {WOS:000863323200005},
doi = {10.1016/j.jbc.2022.102317},
url = {https://bib-pubdb1.desy.de/record/486058},
}