{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T17:12:02Z","timestamp":1786727522955,"version":"build-2736575974"},"reference-count":69,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2024,11,19]],"date-time":"2024-11-19T00:00:00Z","timestamp":1731974400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Institutes of Health","award":["HD102623"],"award-info":[{"award-number":["HD102623"]}]},{"name":"National Institutes of Health","award":["#R344-2020-1023"],"award-info":[{"award-number":["#R344-2020-1023"]}]},{"name":"National Institutes of Health","award":["24K01975"],"award-info":[{"award-number":["24K01975"]}]},{"name":"National Institutes of Health","award":["JPMJCR22E4"],"award-info":[{"award-number":["JPMJCR22E4"]}]},{"name":"National Institutes of Health","award":["JP24ama121001"],"award-info":[{"award-number":["JP24ama121001"]}]},{"name":"Lundbeck Ascending Investigator","award":["HD102623"],"award-info":[{"award-number":["HD102623"]}]},{"name":"Lundbeck Ascending Investigator","award":["#R344-2020-1023"],"award-info":[{"award-number":["#R344-2020-1023"]}]},{"name":"Lundbeck Ascending Investigator","award":["24K01975"],"award-info":[{"award-number":["24K01975"]}]},{"name":"Lundbeck Ascending Investigator","award":["JPMJCR22E4"],"award-info":[{"award-number":["JPMJCR22E4"]}]},{"name":"Lundbeck Ascending Investigator","award":["JP24ama121001"],"award-info":[{"award-number":["JP24ama121001"]}]},{"name":"Grant-in-Aid for Scientific Research","award":["HD102623"],"award-info":[{"award-number":["HD102623"]}]},{"name":"Grant-in-Aid for Scientific Research","award":["#R344-2020-1023"],"award-info":[{"award-number":["#R344-2020-1023"]}]},{"name":"Grant-in-Aid for Scientific Research","award":["24K01975"],"award-info":[{"award-number":["24K01975"]}]},{"name":"Grant-in-Aid for Scientific Research","award":["JPMJCR22E4"],"award-info":[{"award-number":["JPMJCR22E4"]}]},{"name":"Grant-in-Aid for Scientific Research","award":["JP24ama121001"],"award-info":[{"award-number":["JP24ama121001"]}]},{"name":"Daiich Sankyo Foundation of Life Science, JST CREST","award":["HD102623"],"award-info":[{"award-number":["HD102623"]}]},{"name":"Daiich Sankyo Foundation of Life Science, JST CREST","award":["#R344-2020-1023"],"award-info":[{"award-number":["#R344-2020-1023"]}]},{"name":"Daiich Sankyo Foundation of Life Science, JST CREST","award":["24K01975"],"award-info":[{"award-number":["24K01975"]}]},{"name":"Daiich Sankyo Foundation of Life Science, JST CREST","award":["JPMJCR22E4"],"award-info":[{"award-number":["JPMJCR22E4"]}]},{"name":"Daiich Sankyo Foundation of Life Science, JST CREST","award":["JP24ama121001"],"award-info":[{"award-number":["JP24ama121001"]}]},{"name":"AMED","award":["HD102623"],"award-info":[{"award-number":["HD102623"]}]},{"name":"AMED","award":["#R344-2020-1023"],"award-info":[{"award-number":["#R344-2020-1023"]}]},{"name":"AMED","award":["24K01975"],"award-info":[{"award-number":["24K01975"]}]},{"name":"AMED","award":["JPMJCR22E4"],"award-info":[{"award-number":["JPMJCR22E4"]}]},{"name":"AMED","award":["JP24ama121001"],"award-info":[{"award-number":["JP24ama121001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Na+,K+-ATPase is the active ion transport system that maintains the electrochemical gradients for Na+ and K+ across the plasma membrane of most animal cells. Na+,K+-ATPase is constituted by the association of two major subunits, a catalytic \u03b1 and a glycosylated \u03b2 subunit, both of which exist as different isoforms (in mammals known as \u03b11, \u03b12, \u03b13, \u03b14, \u03b21, \u03b22 and \u03b23). Na+,K+-ATPase \u03b1 and \u03b2 isoforms assemble in different combinations to produce various isozymes with tissue specific expression and distinct biochemical properties. Na+,K+-ATPase \u03b14\u03b21 is only found in male germ cells of the testis and is mainly expressed in the sperm flagellum, where it plays a critical role in sperm motility and male fertility. Here, we report the molecular structure of Na+,K+-ATPase \u03b14\u03b21 at 2.37 \u00c5 resolution in the ouabain-bound state and in the presence of beryllium fluoride. Overall, Na+,K+-ATPase \u03b14 structure exhibits the basic major domains of a P-Type ATPase, resembling Na+,K+-ATPase \u03b11, but has differences specific to its distinct sequence. Dissimilarities include the site where the inhibitor ouabain binds. Molecular simulations indicate that glycosphingolipids can bind to a putative glycosphingolipid binding site, which could potentially modulate Na+,K+-ATPase \u03b14 activity. This is the first experimental evidence for the structure of Na+,K+-ATPase \u03b14\u03b21. These data provide a template that will aid in better understanding the function Na+,K+-ATPase \u03b14\u03b21 and will be important for the design and development of compounds that can modulate Na+,K+-ATPase \u03b14 activity for the purpose of improving male fertility or to achieve male contraception.<\/jats:p>","DOI":"10.3390\/ijms252212397","type":"journal-article","created":{"date-parts":[[2024,11,19]],"date-time":"2024-11-19T04:53:05Z","timestamp":1731991985000},"page":"12397","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Molecular Structure of the Na+,K+-ATPase \u03b14\u03b21 Isoform in Its Ouabain-Bound Conformation"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2681-5921","authenticated-orcid":false,"given":"Kazuhiro","family":"Abe","sequence":"first","affiliation":[{"name":"Department of Chemistry, Faculty of Science, Hokkaido University, Hokkaido 060-0808, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3566-3483","authenticated-orcid":false,"given":"Jeff","family":"McDermott","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Physiology, The University of Kansas Medical Center, Kansas City, KS 66103, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hridya","family":"Valia Madapally","sequence":"additional","affiliation":[{"name":"PhyLife: Physical Life Science, Department of Physics Chemistry and Pharmacy, University of Southern Denmark, 5230 Odense, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4960-2160","authenticated-orcid":false,"given":"Parthiban","family":"Marimuthu","sequence":"additional","affiliation":[{"name":"Pharmaceutical Science Laboratory (PSL\u2014Pharmacy) and Structural Bioinformatics Laboratory (SBL\u2014Biochemistry), Faculty of Science and Engineering, \u00c5bo Akademi University, FI-20520 Turku, Finland"},{"name":"Center for Global Health Research, Saveetha Medical College, Saveetha Institute of Medical and Technical Sciences, Chennai 602 105, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9014-9380","authenticated-orcid":false,"given":"Chai C.","family":"Gopalasingam","sequence":"additional","affiliation":[{"name":"RIKEN SPring-8 Center, Kouto, Sayo-gun, Hyogo 679-5148, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7265-2804","authenticated-orcid":false,"given":"Christoph","family":"Gerle","sequence":"additional","affiliation":[{"name":"RIKEN SPring-8 Center, Kouto, Sayo-gun, Hyogo 679-5148, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3951-8651","authenticated-orcid":false,"given":"Hideki","family":"Shigematsu","sequence":"additional","affiliation":[{"name":"Japan Synchrotron Radiation Research Institute (JASRI), SPring-8, 1-1-1 Kouto, Sayo, Hyogo 679-5148, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Himanshu","family":"Khandelia","sequence":"additional","affiliation":[{"name":"PhyLife: Physical Life Science, Department of Physics Chemistry and Pharmacy, University of Southern Denmark, 5230 Odense, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7721-9869","authenticated-orcid":false,"given":"Gustavo","family":"Blanco","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Physiology, The University of Kansas Medical Center, Kansas City, KS 66103, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,11,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1146\/annurev.biochem.71.102201.141218","article-title":"Biochemistry of Na,K-ATPase","volume":"71","author":"Kaplan","year":"2002","journal-title":"Annu. 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