{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,18]],"date-time":"2026-01-18T21:24:24Z","timestamp":1768771464080,"version":"3.49.0"},"reference-count":26,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2025,3,17]],"date-time":"2025-03-17T00:00:00Z","timestamp":1742169600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Molecular Microbiology"],"published-print":{"date-parts":[[2025,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>The sodium\/proton\u2010exchanging ATPase of <jats:italic>Plasmodium falciparum<\/jats:italic> malaria parasites, PfATP4, is an emerging drug target. Inhibition results in detrimental cell swelling due to cytosolic accumulation of sodium and alkalization. PfATP4 is a sodium\u2010releasing type II P\u2010type ATPase restricted to apicomplexan parasites. Experimental data on structure\u2013function relationships of the isolated protein are absent. Here, we produced and purified the soluble catalytic domain of PfATP4 and evaluated kinetic properties by in\u00a0vitro phosphate colorimetry. The protein exhibited Mg<jats:sup>2+<\/jats:sup>\u2010dependent ATPase activity at the same order of magnitude as the native cellular PfATP4 and was insensitive to the presence of sodium. AlphaFold 3\u2010based structure and ATP\/Mg<jats:sup>2+<\/jats:sup> interaction predictions identified key residues of the nucleotide binding domain (Lys619, Lys652, Arg703). Replacement of the lysines by methionine decreased the enzymatic activity to one quarter. Individual mutation of the putative Mg<jats:sup>2+<\/jats:sup>\u2010coordinating Asp865 of the phosphorylation domain was tolerated, while a joint replacement with Asp869 decreased ATPase again to one quarter. Mutation of the putative \u03b3\u2010phosphate receiving Asp451 maintained the rate of P<jats:sub>i<\/jats:sub> release. Our data attribute typical functional roles for P\u2010type ATPases to the basic and acidic residues of the soluble PfATP4 catalytic domain and show that its ATP hydrolysis is independent of phosphorylation of Asp451.<\/jats:p>","DOI":"10.1111\/mmi.15358","type":"journal-article","created":{"date-parts":[[2025,3,17]],"date-time":"2025-03-17T05:54:36Z","timestamp":1742190876000},"page":"479-486","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["A Soluble Expression Construct of the Isolated Catalytic Domain of <i>Plasmodium falciparum<\/i><scp>ATP4<\/scp> Exhibits <scp>ATPase<\/scp> Activity Independent of a \u03b3\u2010Phosphate Receiving Aspartate"],"prefix":"10.1111","volume":"123","author":[{"given":"Timo","family":"Beyer","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical and Medicinal Chemistry Christian\u2010Albrechts\u2010University of Kiel  Kiel Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jesko","family":"Caliebe","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical and Medicinal Chemistry Christian\u2010Albrechts\u2010University of Kiel  Kiel Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lara","family":"K\u00e4hler","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical and Medicinal Chemistry Christian\u2010Albrechts\u2010University of Kiel  Kiel Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5912-6626","authenticated-orcid":false,"given":"Eric","family":"Beitz","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical and Medicinal Chemistry Christian\u2010Albrechts\u2010University of Kiel  Kiel 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