{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,10]],"date-time":"2026-06-10T16:59:46Z","timestamp":1781110786726,"version":"3.54.1"},"reference-count":52,"publisher":"National Academy of Sciences","issue":"2","license":[{"start":{"date-parts":[[2025,1,8]],"date-time":"2025-01-08T00:00:00Z","timestamp":1736294400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000060","name":"HHS | NIH | National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01AI132508"],"award-info":[{"award-number":["R01AI132508"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"HHS | NIH | National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01AI154499"],"award-info":[{"award-number":["R01AI154499"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Anton 2 Award","award":["MCB210021P"],"award-info":[{"award-number":["MCB210021P"]}]},{"DOI":"10.13039\/100000057","name":"HHS | NIH | National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["R01GM116961"],"award-info":[{"award-number":["R01GM116961"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2025,1,14]]},"abstract":"<jats:p>\n                    Among new antimalarials discovered over the past decade are multiple chemical scaffolds that target\n                    <jats:italic>Plasmodium falciparum<\/jats:italic>\n                    P-type ATPase (\n                    <jats:italic>Pf<\/jats:italic>\n                    ATP4). This essential protein is a Na\n                    <jats:sup>+<\/jats:sup>\n                    pump responsible for the maintenance of Na\n                    <jats:sup>+<\/jats:sup>\n                    homeostasis.\n                    <jats:italic>Pf<\/jats:italic>\n                    ATP4 belongs to the type 2D subfamily of P-type ATPases, for which no structures have been determined. To gain better insight into the structure\/function relationship of this validated drug target, we generated a homology model of\n                    <jats:italic>Pf<\/jats:italic>\n                    ATP4 based on sarco\/endoplasmic reticulum Ca\n                    <jats:sup>2+<\/jats:sup>\n                    ATPase, a P2A-type ATPase, and refined the model using molecular dynamics in its explicit membrane environment. This model predicted several residues in\n                    <jats:italic>Pf<\/jats:italic>\n                    ATP4 critical for its function, as well as those that impart resistance to various\n                    <jats:italic>Pf<\/jats:italic>\n                    ATP4 inhibitors. To validate our model, we developed a genetic system involving merodiploid states of\n                    <jats:italic>Pf<\/jats:italic>\n                    ATP4 in which the endogenous gene was conditionally expressed, and the second allele was mutated to assess its effect on the parasite. Our model predicted residues involved in Na\n                    <jats:sup>+<\/jats:sup>\n                    coordination as well as the phosphorylation cycle of\n                    <jats:italic>Pf<\/jats:italic>\n                    ATP4. Phenotypic characterization of these mutants involved assessment of parasite growth, localization of mutated\n                    <jats:italic>Pf<\/jats:italic>\n                    ATP4, response to treatment with known\n                    <jats:italic>Pf<\/jats:italic>\n                    ATP4 inhibitors, and evaluation of the downstream consequences of Na\n                    <jats:sup>+<\/jats:sup>\n                    influx. Our results were consistent with modeled predictions of the essentiality of the critical residues. Additionally, our approach confirmed the phenotypic consequences of resistance-associated mutations as well as a potential structural basis for the fitness cost associated with some mutations. Taken together, our approach provides a means to explore the structure\/function relationship of essential genes in haploid organisms.\n                  <\/jats:p>","DOI":"10.1073\/pnas.2403689122","type":"journal-article","created":{"date-parts":[[2025,1,8]],"date-time":"2025-01-08T11:25:57Z","timestamp":1736335557000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":4,"title":["Mutational analysis of an antimalarial drug target,\n                    <i>Pf<\/i>\n                    ATP4"],"prefix":"10.1073","volume":"122","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-4290-9795","authenticated-orcid":false,"given":"Swaksha","family":"Rachuri","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, Center for Molecular Parasitology, Drexel University College of Medicine","place":["Philadelphia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Binod","family":"Nepal","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Center for Molecular Parasitology, Drexel University College of Medicine","place":["Philadelphia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4035-2978","authenticated-orcid":false,"given":"Anurag","family":"Shukla","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Center for Molecular Parasitology, Drexel University College of Medicine","place":["Philadelphia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aarti","family":"Ramanathan","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Center for Molecular Parasitology, Drexel University College of 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College of Medicine","place":["Philadelphia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lawrence W.","family":"Bergman","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Center for Molecular Parasitology, Drexel University College of Medicine","place":["Philadelphia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3532-8669","authenticated-orcid":false,"given":"Sandhya","family":"Kortagere","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Center for Molecular Parasitology, Drexel University College of Medicine","place":["Philadelphia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1063-5571","authenticated-orcid":false,"given":"Akhil B.","family":"Vaidya","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Center for Molecular Parasitology, Drexel 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