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==Cryo-EM structure of the gastric proton pump K791S/E820D/Y340N/E936V/Y799W mutant in K+-occluded (K+)E2-AlF state== | |||
<StructureSection load='7et1' size='340' side='right'caption='[[7et1]], [[Resolution|resolution]] 2.60Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[7et1]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Sus_scrofa Sus scrofa]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7ET1 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7ET1 FirstGlance]. <br> | |||
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">Electron Microscopy, [[Resolution|Resolution]] 2.6Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ALF:TETRAFLUOROALUMINATE+ION'>ALF</scene>, <scene name='pdbligand=CLR:CHOLESTEROL'>CLR</scene>, <scene name='pdbligand=K:POTASSIUM+ION'>K</scene>, <scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene>, <scene name='pdbligand=NAG:N-ACETYL-D-GLUCOSAMINE'>NAG</scene>, <scene name='pdbligand=PCW:1,2-DIOLEOYL-SN-GLYCERO-3-PHOSPHOCHOLINE'>PCW</scene></td></tr> | |||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=7et1 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7et1 OCA], [https://pdbe.org/7et1 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7et1 RCSB], [https://www.ebi.ac.uk/pdbsum/7et1 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7et1 ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[https://www.uniprot.org/uniprot/ATP4A_PIG ATP4A_PIG] Catalyzes the hydrolysis of ATP coupled with the exchange of H(+) and K(+) ions across the plasma membrane. Responsible for acid production in the stomach. | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
The gastric H(+),K(+)-ATPase mediates electroneutral exchange of 1H(+)/1K(+) per ATP hydrolysed across the membrane. Previous structural analysis of the K(+)-occluded E2-P transition state of H(+),K(+)-ATPase showed a single bound K(+) at cation-binding site II, in marked contrast to the two K(+) ions occluded at sites I and II of the closely-related Na(+),K(+)-ATPase which mediates electrogenic 3Na(+)/2K(+) translocation across the membrane. The molecular basis of the different K(+) stoichiometry between these K(+)-counter-transporting pumps is elusive. We show a series of crystal structures and a cryo-EM structure of H(+),K(+)-ATPase mutants with changes in the vicinity of site I, based on the structure of the sodium pump. Our step-wise and tailored construction of the mutants finally gave a two-K(+) bound H(+),K(+)-ATPase, achieved by five mutations, including amino acids directly coordinating K(+) (Lys791Ser, Glu820Asp), indirectly contributing to cation-binding site formation (Tyr340Asn, Glu936Val), and allosterically stabilizing K(+)-occluded conformation (Tyr799Trp). This quintuple mutant in the K(+)-occluded E2-P state unambiguously shows two separate densities at the cation-binding site in its 2.6 A resolution cryo-EM structure. These results offer new insights into how two closely-related cation pumps specify the number of K(+) accommodated at their cation-binding site. | |||
Gastric proton pump with two occluded K(+) engineered with sodium pump-mimetic mutations.,Abe K, Yamamoto K, Irie K, Nishizawa T, Oshima A Nat Commun. 2021 Sep 29;12(1):5709. doi: 10.1038/s41467-021-26024-1. PMID:34588453<ref>PMID:34588453</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
[[Category: Abe | <div class="pdbe-citations 7et1" style="background-color:#fffaf0;"></div> | ||
==See Also== | |||
*[[ATPase 3D structures|ATPase 3D structures]] | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Large Structures]] | |||
[[Category: Sus scrofa]] | |||
[[Category: Abe K]] | |||
Latest revision as of 11:42, 17 October 2024
Cryo-EM structure of the gastric proton pump K791S/E820D/Y340N/E936V/Y799W mutant in K+-occluded (K+)E2-AlF stateCryo-EM structure of the gastric proton pump K791S/E820D/Y340N/E936V/Y799W mutant in K+-occluded (K+)E2-AlF state
Structural highlights
FunctionATP4A_PIG Catalyzes the hydrolysis of ATP coupled with the exchange of H(+) and K(+) ions across the plasma membrane. Responsible for acid production in the stomach. Publication Abstract from PubMedThe gastric H(+),K(+)-ATPase mediates electroneutral exchange of 1H(+)/1K(+) per ATP hydrolysed across the membrane. Previous structural analysis of the K(+)-occluded E2-P transition state of H(+),K(+)-ATPase showed a single bound K(+) at cation-binding site II, in marked contrast to the two K(+) ions occluded at sites I and II of the closely-related Na(+),K(+)-ATPase which mediates electrogenic 3Na(+)/2K(+) translocation across the membrane. The molecular basis of the different K(+) stoichiometry between these K(+)-counter-transporting pumps is elusive. We show a series of crystal structures and a cryo-EM structure of H(+),K(+)-ATPase mutants with changes in the vicinity of site I, based on the structure of the sodium pump. Our step-wise and tailored construction of the mutants finally gave a two-K(+) bound H(+),K(+)-ATPase, achieved by five mutations, including amino acids directly coordinating K(+) (Lys791Ser, Glu820Asp), indirectly contributing to cation-binding site formation (Tyr340Asn, Glu936Val), and allosterically stabilizing K(+)-occluded conformation (Tyr799Trp). This quintuple mutant in the K(+)-occluded E2-P state unambiguously shows two separate densities at the cation-binding site in its 2.6 A resolution cryo-EM structure. These results offer new insights into how two closely-related cation pumps specify the number of K(+) accommodated at their cation-binding site. Gastric proton pump with two occluded K(+) engineered with sodium pump-mimetic mutations.,Abe K, Yamamoto K, Irie K, Nishizawa T, Oshima A Nat Commun. 2021 Sep 29;12(1):5709. doi: 10.1038/s41467-021-26024-1. PMID:34588453[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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