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| 001 | 486058 | ||
| 005 | 20250724152116.0 | ||
| 024 | 7 | _ | |a 10.1016/j.jbc.2022.102317 |2 doi |
| 024 | 7 | _ | |a 0021-9258 |2 ISSN |
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| 100 | 1 | _ | |a Fruergaard, Marlene U. |b 0 |
| 245 | _ | _ | |a The Na$^+$,K$^+$-ATPase in complex with beryllium fluoride mimics an ATPase phosphorylated state |
| 260 | _ | _ | |a Bethesda, MD. |c 2022 |b American Soc. for Biochemistry and Molecular Biology |
| 264 | _ | 1 | |3 print |2 Crossref |b Elsevier BV |c 2022-09-01 |
| 264 | _ | 1 | |3 print |2 Crossref |b Elsevier BV |c 2022-09-01 |
| 336 | 7 | _ | |a article |2 DRIVER |
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| 336 | 7 | _ | |a Journal Article |b journal |m journal |0 PUB:(DE-HGF)16 |s 1669380876_18186 |2 PUB:(DE-HGF) |
| 336 | 7 | _ | |a ARTICLE |2 BibTeX |
| 336 | 7 | _ | |a JOURNAL_ARTICLE |2 ORCID |
| 336 | 7 | _ | |a Journal Article |0 0 |2 EndNote |
| 500 | _ | _ | |a ISSN 0021-9258 not unique: **2 hits**. |
| 520 | _ | _ | |a 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. |
| 536 | _ | _ | |a 6G3 - PETRA III (DESY) (POF4-6G3) |0 G:(DE-HGF)POF4-6G3 |c POF4-6G3 |f POF IV |x 0 |
| 542 | _ | _ | |i 2022-09-01 |2 Crossref |u https://www.elsevier.com/tdm/userlicense/1.0/ |
| 542 | _ | _ | |i 2022-07-28 |2 Crossref |u http://creativecommons.org/licenses/by/4.0/ |
| 588 | _ | _ | |a Dataset connected to CrossRef, Journals: bib-pubdb1.desy.de |
| 693 | _ | _ | |a PETRA III |f PETRA Beamline P14 |1 EXP:(DE-H253)PETRAIII-20150101 |0 EXP:(DE-H253)P-P14-20150101 |6 EXP:(DE-H253)P-P14-20150101 |x 0 |
| 693 | _ | _ | |a PETRA III |f PETRA Beamline P13 |1 EXP:(DE-H253)PETRAIII-20150101 |0 EXP:(DE-H253)P-P13-20150101 |6 EXP:(DE-H253)P-P13-20150101 |x 1 |
| 700 | 1 | _ | |a Dach, Ingrid |b 1 |
| 700 | 1 | _ | |a Andersen, Jacob L. |b 2 |
| 700 | 1 | _ | |a Ozol, Mette |b 3 |
| 700 | 1 | _ | |a Shahsavar, Azadeh |b 4 |
| 700 | 1 | _ | |a Quistgaard, Esben M. |b 5 |
| 700 | 1 | _ | |a Poulsen, Hanne |b 6 |
| 700 | 1 | _ | |a Fedosova, Natalya U. |0 0000-0002-1706-6108 |b 7 |e Corresponding author |
| 700 | 1 | _ | |a Nissen, Poul |0 P:(DE-H253)PIP1087253 |b 8 |e Corresponding author |
| 773 | 1 | 8 | |a 10.1016/j.jbc.2022.102317 |b Elsevier BV |d 2022-09-01 |n 9 |p 102317 |3 journal-article |2 Crossref |t Journal of Biological Chemistry |v 298 |y 2022 |x 0021-9258 |
| 773 | _ | _ | |a 10.1016/j.jbc.2022.102317 |g Vol. 298, no. 9, p. 102317 - |0 PERI:(DE-600)1474604-9 |n 9 |p 102317 |t The journal of biological chemistry |v 298 |y 2022 |x 0021-9258 |
| 856 | 4 | _ | |u https://www.sciencedirect.com/science/article/pii/S0021925822007591?via%3Dihub |
| 856 | 4 | _ | |u https://bib-pubdb1.desy.de/record/486058/files/The%20Na%2B%2CK%2B-ATPase%20in%20complex%20with%20beryllium%20fluoride%20mimics%20an%20ATPase%20phosphorylated%20state.pdf |y OpenAccess |
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