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Inhibiting B\n                    <jats:sup>0<\/jats:sup>\n                    AT1 enhances Phe excretion, offering a therapeutic strategy for phenylketonuria (PKU). Using cryo-EM, we determined human B\n                    <jats:sup>0<\/jats:sup>\n                    AT1 structures in outward- and inward-open states, revealing an allosteric pocket ~17\u2009\u00c5 from the substrate site that is present in the outward-open conformation and has been previously reported. Structure-guided inhibitor design targeting this pocket produced a potent B\n                    <jats:sup>0<\/jats:sup>\n                    AT1 inhibitor that locks the transporter in an outward-occluded state and blocks transport. The higher-resolution structures reveal detailed interactions at the binding site, including water-mediated coordination and conformational changes around Leu52. This inhibitor exhibited submicromolar IC\n                    <jats:sub>50<\/jats:sub>\n                    against both human and mouse B\n                    <jats:sup>0<\/jats:sup>\n                    AT1, and oral administration in PKU model mice increased urinary Phe and reduced plasma Phe levels. These findings provide structural insight into allosteric inhibition of B\n                    <jats:sup>0<\/jats:sup>\n                    AT1 and establish a framework for the rational optimization of inhibitors targeting conformationally dynamic allosteric sites in SLC6-family transporters.\n                  <\/jats:p>","DOI":"10.1038\/s42003-026-10535-y","type":"journal-article","created":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T17:07:09Z","timestamp":1782407229000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Structure-guided development of a potent human B0AT1 inhibitor effective in a mouse model of phenylketonuria"],"prefix":"10.1038","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-9340-380X","authenticated-orcid":false,"given":"Takuya","family":"Imazu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomoya","family":"Akashi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4340-2937","authenticated-orcid":false,"given":"Masahiro","family":"Hiraizumi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yosuke","family":"Inui","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wataru","family":"Sasaki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tsuyoshi","family":"Takahashi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hidenori","family":"Todoroki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taichi","family":"Kumanomidou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazunori","family":"Yamada","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Norie","family":"Fujikawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiromi","family":"Hisano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hidetsugu","family":"Asada","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6186-6647","authenticated-orcid":false,"given":"Tsukasa","family":"Kusakizako","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7463-8398","authenticated-orcid":false,"given":"Tomohiro","family":"Nishizawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"So","family":"Iwata","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1813-7008","authenticated-orcid":false,"given":"Osamu","family":"Nureki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0021-7296","authenticated-orcid":false,"given":"Ikuko","family":"Miyaguchi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,6,25]]},"reference":[{"key":"10535_CR1","doi-asserted-by":"publisher","first-page":"234","DOI":"10.1016\/j.ajhg.2020.06.006","volume":"107","author":"A Hillert","year":"2020","unstructured":"Hillert, A. et al. 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I.M. was a former employee of Mitsubishi Tanabe Pharma Co., Ltd., and is currently employed by Prism BioLab, Inc.; T.A. by Chugai Pharmaceutical Co., Ltd.; and M.H. by the University of Tokyo. These current affiliations are unrelated to the present research. The other authors declare no competing interests.","order":1,"name":"Ethics","label":"Competing interests","group":{"name":"EthicsHeading","label":"Ethics"}}],"article-number":"1350"}}