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We have recently uncovered that the signaling molecule diadenosine tetraphosphate (Ap4A) acts as a crucial regulator of\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    virulence. Specifically, deletion of the Ap4A\u2010degrading enzyme, diadenosine tetraphosphatase (ApaH), dramatically reduces the expression of key virulence factors. The structural and molecular properties of\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    ApaH (\n                    <jats:italic>Pa<\/jats:italic>\n                    ApaH) remain uncharacterized. Here, we present an integrated biochemical, structural, computational, and phenotypic characterization of\n                    <jats:italic>Pa<\/jats:italic>\n                    ApaH. We define the molecular determinants of its manganese\u2010dependent Ap4A hydrolytic mechanism and establish a direct functional link between\n                    <jats:italic>Pa<\/jats:italic>\n                    ApaH catalytic activity and virulence phenotypes\n                    <jats:italic>in vivo<\/jats:italic>\n                    . Together with the absence of ApaH homologs in eukaryotes, these findings place\n                    <jats:italic>Pa<\/jats:italic>\n                    ApaH as an attractive and selective target for antivirulence therapeutic strategies against\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    infections.\n                  <\/jats:p>","DOI":"10.1002\/pro.70781","type":"journal-article","created":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T15:00:52Z","timestamp":1787670052000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Structural and functional insights into\n                    <i>Pseudomonas aeruginosa<\/i>\n                    <scp>ApaH<\/scp>\n                    , a diadenosine tetraphosphatase crucial for bacterial virulence"],"prefix":"10.1002","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-1763-0331","authenticated-orcid":false,"given":"Gianluca","family":"Pistoia","sequence":"first","affiliation":[{"name":"Department of Biochemical Sciences \u201cA. Rossi Fanelli\u201d Sapienza University of Rome  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4292-2615","authenticated-orcid":false,"given":"Matteo","family":"Cervoni","sequence":"additional","affiliation":[{"name":"Department of Science University Roma Tre  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-7793-3577","authenticated-orcid":false,"given":"Flavia","family":"Catalano","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology and Pathology National Research Council  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1377-8527","authenticated-orcid":false,"given":"Francesca","family":"Troilo","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology and Pathology National Research Council  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-3641-310X","authenticated-orcid":false,"given":"Francesca","family":"Guidi","sequence":"additional","affiliation":[{"name":"Department of Science University Roma Tre  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-2569-0709","authenticated-orcid":false,"given":"Eleonora","family":"Comparini","sequence":"additional","affiliation":[{"name":"Department of Biochemical Sciences \u201cA. Rossi Fanelli\u201d Sapienza University of Rome  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppina","family":"Mignogna","sequence":"additional","affiliation":[{"name":"Department of Biochemical Sciences \u201cA. Rossi Fanelli\u201d Sapienza University of Rome  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlo","family":"Travaglini\u2010Allocatelli","sequence":"additional","affiliation":[{"name":"Department of Biochemical Sciences \u201cA. Rossi Fanelli\u201d Sapienza University of Rome  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5301-0681","authenticated-orcid":false,"given":"Alessandro","family":"Giuffr\u00e8","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology and Pathology National Research Council  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7940-8206","authenticated-orcid":false,"given":"Antonio","family":"Coluccia","sequence":"additional","affiliation":[{"name":"Department of Drug Chemistry and Technologies Sapienza University of Rome  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5080-5665","authenticated-orcid":false,"given":"Francesco","family":"Imperi","sequence":"additional","affiliation":[{"name":"Department of Science University Roma Tre  Rome Italy"},{"name":"LIFE Institute for Research and Health Care Santa Lucia IRCCS  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5921-7365","authenticated-orcid":false,"given":"Adele","family":"Di Matteo","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology and Pathology National Research Council  Rome Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0802-1370","authenticated-orcid":false,"given":"Giorgio","family":"Giardina","sequence":"additional","affiliation":[{"name":"Department of Biochemical Sciences \u201cA. 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