{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T23:37:47Z","timestamp":1768520267446,"version":"3.49.0"},"posted":{"date-parts":[[2025,1,11]]},"group-title":"Microbiology","reference-count":66,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,1,11]],"date-time":"2025-01-11T00:00:00Z","timestamp":1736553600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,1,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  <jats:italic>Candida albicans<\/jats:italic>\n                  is a growing health concern as the leading causal agent of systemic candidiasis, a life-threatening fungal infection with a mortality rate of \u223c40% despite best available therapy. Yck2, a fungal casein kinase 1 (CK1) family member, is the cellular target of inhibitors YK-I-02 (YK) and MN-I-157 (MN). Here, multiplexed inhibitor beads paired with mass spectrometry (MIB\/MS) employing ATP-competitive kinase inhibitors were used to define the selectivity of these Yck2 inhibitors across the global\n                  <jats:italic>C. albicans<\/jats:italic>\n                  proteome. The MIB matrix captured 89% of the known and predicted\n                  <jats:italic>C. albicans<\/jats:italic>\n                  protein kinases present in cell lysate. In MIB\/MS competition assays, YK and MN demonstrated exquisite selectivity across the\n                  <jats:italic>C. albicans<\/jats:italic>\n                  fungal kinome with target engagement of only three CK1 homologs (Yck2, Yck22, and Hrr25) and a homolog of human p38\u03b1 (Hog1). Additional chemoproteomics using a custom MN-kinobead identified only one additional\n                  <jats:italic>C. albicans<\/jats:italic>\n                  protein, confirming its remarkable fungal proteome-wide selectivity. To identify new Yck2 inhibitors with selectivity over Hog1, thirteen human CK1 kinase inhibitors were profiled for fungal kinase-binding activity using MIB\/MS competition assays and in-cell NanoBRET target engagement assays. A new chemotype of family-selective Yck2 inhibitors with antifungal activity was identified. Together, these findings expand the application of MIB\/MS proteomic profiling for non-human kinomes and demonstrate its utility in the discovery and development of selective inhibitors of fungal kinases with potential antimicrobial activity.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>GRAPHICAL ABSTRACT<\/jats:title>\n                  <jats:fig id=\"ufig1\" position=\"float\" orientation=\"portrait\" fig-type=\"figure\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"632200v1_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:fig>\n                <\/jats:sec>","DOI":"10.1101\/2025.01.10.632200","type":"posted-content","created":{"date-parts":[[2025,1,11]],"date-time":"2025-01-11T04:50:18Z","timestamp":1736571018000},"source":"Crossref","is-referenced-by-count":0,"title":["Chemoproteomic Profiling of\n                  <i>C. albicans<\/i>\n                  for Characterization of Anti-fungal Kinase Inhibitors"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-2475-7366","authenticated-orcid":false,"given":"David J.","family":"Shirley","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9093-2244","authenticated-orcid":false,"given":"Meganathan","family":"Nandakumar","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aurora","family":"Cabrera","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bonnie","family":"Yiu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-3371-5877","authenticated-orcid":false,"given":"Emily","family":"Puumala","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4198-8214","authenticated-orcid":false,"given":"Zhongle","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2821-9269","authenticated-orcid":false,"given":"Nicole","family":"Robbins","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6445-5766","authenticated-orcid":false,"given":"Luke","family":"Whitesell","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2189-0420","authenticated-orcid":false,"given":"Jeffrey L.","family":"Smith","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3004-0148","authenticated-orcid":false,"given":"Scott P.","family":"Lyons","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0151-4498","authenticated-orcid":false,"given":"Angie L.","family":"Mordant","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4496-7312","authenticated-orcid":false,"given":"Laura E.","family":"Herring","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4736-9855","authenticated-orcid":false,"given":"Lee M.","family":"Graves","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rafael M.","family":"Cou\u00f1ago","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5973-5798","authenticated-orcid":false,"given":"David H.","family":"Drewry","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5797-0110","authenticated-orcid":false,"given":"Leah E.","family":"Cowen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4181-8223","authenticated-orcid":false,"given":"Timothy M.","family":"Willson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025011303050701000_2025.01.10.632200v1.1","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00449-20"},{"key":"2025011303050701000_2025.01.10.632200v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(23)00692-8"},{"key":"2025011303050701000_2025.01.10.632200v1.3","doi-asserted-by":"publisher","DOI":"10.1126\/science.1222236"},{"key":"2025011303050701000_2025.01.10.632200v1.4","unstructured":"W.H.O. 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