{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T17:03:24Z","timestamp":1790269404077,"version":"4.1.0"},"reference-count":0,"publisher":"Oxford University Press (OUP)","license":[{"start":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T00:00:00Z","timestamp":1790208000000},"content-version":"am","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Golgi-resident 3\u2032-phosphoadenosine-5\u2032-phosphate (PAP) phosphatase BPNT2\/gPAPP plays an essential role in maintaining sulfation homeostasis, but its biochemical characterization has been limited by the difficulty of in vitro analyses. Here, we established a stable expression and activity assay system using a Trigger factor-fused BPNT2 construct. Using this system, we performed AlphaFold 3-guided mutational analyses. The predicted model and mutational analyses showed that several acidic residues are required for efficient PAP phosphatase activity, likely through their contribution to Mg\u00b2\u207a coordination. In addition, T177 was suggested to help maintain an active-site geometry that could accommodate a catalytic water molecule. Conservation analysis and lithium inhibition supported the idea that BPNT2 shares a conserved Mg\u00b2\u207a-dependent catalytic framework with related PAP phosphatases. Disease-associated variants D175N and T181P showed substantially reduced activity, possibly due to impaired Mg2+ coordination and local structural distortion. These findings provide a biochemical framework for understanding BPNT2 function in Golgi sulfation regulation.<\/jats:p>","DOI":"10.1093\/bbb\/zbag144","type":"journal-article","created":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T16:22:17Z","timestamp":1790266937000},"source":"Crossref","is-referenced-by-count":0,"title":["Biochemical analysis of catalytic and disease-associated residues in mouse Golgi-resident 3\u2032-phosphoadenosine-5\u2032-phosphate (PAP) phosphatase BPNT2\/gPAPP"],"prefix":"10.1093","author":[{"given":"Ryota","family":"Urushihara","sequence":"first","affiliation":[{"name":"Laboratory of Biophysical Chemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University , 744 Motooka, Nishi-ku, Fukuoka 819-0395 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasuhisa","family":"Maruno","sequence":"additional","affiliation":[{"name":"Laboratory of Biophysical Chemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University , 744 Motooka, Nishi-ku, Fukuoka 819-0395 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saki","family":"Maruoka","sequence":"additional","affiliation":[{"name":"Laboratory of Biophysical Chemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University , 744 Motooka, Nishi-ku, Fukuoka 819-0395 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moe","family":"Tsuruta","sequence":"additional","affiliation":[{"name":"Laboratory of Biophysical Chemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University , 744 Motooka, Nishi-ku, Fukuoka 819-0395 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryotaro","family":"Matsuzaki","sequence":"additional","affiliation":[{"name":"Laboratory of Biophysical Chemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University , 744 Motooka, Nishi-ku, Fukuoka 819-0395 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Etsuko","family":"Nishimoto","sequence":"additional","affiliation":[{"name":"Laboratory of Biophysical Chemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University , 744 Motooka, Nishi-ku, Fukuoka 819-0395 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1555-5394","authenticated-orcid":false,"given":"Takamasa","family":"Teramoto","sequence":"additional","affiliation":[{"name":"Laboratory of Biophysical Chemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University , 744 Motooka, Nishi-ku, Fukuoka 819-0395 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0781-2074","authenticated-orcid":false,"given":"Yoshimitsu","family":"Kakuta","sequence":"additional","affiliation":[{"name":"Laboratory of Biophysical Chemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Kyushu University , 744 Motooka, Nishi-ku, Fukuoka 819-0395 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,9,24]]},"container-title":["Bioscience, Biotechnology, and Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/bbb\/advance-article-pdf\/doi\/10.1093\/bbb\/zbag144\/71382477\/zbag144.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/bbb\/advance-article-pdf\/doi\/10.1093\/bbb\/zbag144\/71382477\/zbag144.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T16:22:17Z","timestamp":1790266937000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/bbb\/advance-article\/doi\/10.1093\/bbb\/zbag144\/8833854"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,24]]},"references-count":0,"URL":"https:\/\/doi.org\/10.1093\/bbb\/zbag144","relation":{},"ISSN":["1347-6947"],"issn-type":[{"value":"1347-6947","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,24]]},"article-number":"zbag144"}}