{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,1]],"date-time":"2026-08-01T10:51:48Z","timestamp":1785581508674,"version":"3.56.0"},"reference-count":45,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2026,5,2]],"date-time":"2026-05-02T00:00:00Z","timestamp":1777680000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,5,2]],"date-time":"2026-05-02T00:00:00Z","timestamp":1777680000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["278002225"],"award-info":[{"award-number":["278002225"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2026,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Early\u2010stage drug discovery relies on the availability of stable protein for reliable biophysical characterization of ligand binding. However, many\n                    <jats:italic>Plasmodium falciparum<\/jats:italic>\n                    proteins are challenging to produce in heterologous systems, which limits their experimental utility. To address this, we tested whether ProteinMPNN\u2010guided sequence design could generate stabilized surrogate constructs that retain wild\u2010type\u2010like structure and binding thermodynamics. Designs were generated with constraints to maintain conserved and binding\u2010site residues for three therapeutically relevant targets:\n                    <jats:italic>Pf<\/jats:italic>\n                    BDP1\u2010BRD,\n                    <jats:italic>Pf<\/jats:italic>\n                    BDP4\u2010BRD, and\n                    <jats:italic>Pf<\/jats:italic>\n                    K13\u2010KREP. The resulting constructs showed markedly increased thermal stability. Using\n                    <jats:italic>Pf<\/jats:italic>\n                    BDP1\u2010BRD as a benchmark, isothermal titration calorimetry confirmed that the stabilized variants retained wild\u2010type\u2010like binding thermodynamics with a known ligand. Extending this approach to other targets, a\n                    <jats:italic>Pf<\/jats:italic>\n                    K13\u2010KREP construct led to an apo structure with a binding pocket closely matching the wild type. For\n                    <jats:italic>Pf<\/jats:italic>\n                    BDP4\u2010BRD, virtual screening against a previously reported wild\u2010type crystal structure identified putative binders, while a stabilized surrogate for this otherwise unstable target enabled their experimental validation and the determination of a 1.25\u2009\u00c5 co\u2010crystal structure with a newly identified inhibitor. Our findings demonstrate that computationally stabilized surrogates are practical and effective tools for robust biophysics and structure\u2010enabled drug discovery against otherwise challenging malaria proteins.\n                  <\/jats:p>","DOI":"10.1002\/pro.70614","type":"journal-article","created":{"date-parts":[[2026,5,2]],"date-time":"2026-05-02T10:36:21Z","timestamp":1777718181000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Stabilizing\n                    <scp>\n                      <i>Plasmodium falciparum<\/i>\n                    <\/scp>\n                    proteins for small molecule drug discovery"],"prefix":"10.1002","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7594-3810","authenticated-orcid":false,"given":"Marius","family":"Amann","sequence":"first","affiliation":[{"name":"Institut f\u00fcr Pharmazeutische Wissenschaften Albert\u2010Ludwigs\u2010Universit\u00e4t Freiburg  Freiburg i. Br Germany"},{"name":"Institut f\u00fcr Biochemie Albert\u2010Ludwigs\u2010Universit\u00e4t Freiburg  Freiburg i. Br Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-5014-3921","authenticated-orcid":false,"given":"Tim","family":"Str\u00e4\u00dfer","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Biochemie Albert\u2010Ludwigs\u2010Universit\u00e4t Freiburg  Freiburg i. Br Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8722-2893","authenticated-orcid":false,"given":"Oliver","family":"Einsle","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Biochemie Albert\u2010Ludwigs\u2010Universit\u00e4t Freiburg  Freiburg i. Br Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3744-189X","authenticated-orcid":false,"given":"Stefan","family":"G\u00fcnther","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Pharmazeutische Wissenschaften Albert\u2010Ludwigs\u2010Universit\u00e4t Freiburg  Freiburg i. 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