{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T16:04:21Z","timestamp":1753891461386,"version":"3.41.2"},"reference-count":45,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,7,9]],"date-time":"2024-07-09T00:00:00Z","timestamp":1720483200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003246","name":"Nederlandse Organisatie voor Wetenschappelijk Onderzoek","doi-asserted-by":"publisher","award":["VI.VIDI.192.011"],"award-info":[{"award-number":["VI.VIDI.192.011"]}],"id":[{"id":"10.13039\/501100003246","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001826","name":"ZonMw","doi-asserted-by":"publisher","award":["451001026"],"award-info":[{"award-number":["451001026"]}],"id":[{"id":"10.13039\/501100001826","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000781","name":"European Research Council","doi-asserted-by":"publisher","award":["101087582"],"award-info":[{"award-number":["101087582"]}],"id":[{"id":"10.13039\/501100000781","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Mol. Biosci."],"abstract":"<jats:p>Upon infection of host cells the <jats:italic>Legionella pneumophila<\/jats:italic> bacterium releases a multitude of effector enzymes into the host\u2019s cytoplasm that manipulate cellular host pathways, including the host-ubiquitination pathways. The effectors belonging to the SidE-family are involved in non-canonical phosphoribosyl serine ubiquitination (PR-ubiquitination) of host substrate proteins. This results in the recruitment of ER-remodeling proteins and the formation of a Legionella-containing vacuole which is crucial in the onset of legionnaires disease. PR-ubiquitination is a dynamic process reversed by other <jats:italic>Legionella<\/jats:italic> effectors called Dups. During PR-Ubiquitin phosphodiester hydrolysis Dups form a covalent intermediate with the phosphoribosyl ubiquitylated protein using its active site His67 residue. We envisioned that covalent probes to target <jats:italic>Legionella<\/jats:italic> effectors could be of value to study these effectors and contribute to deciphering the complex biology of <jats:italic>Legionella<\/jats:italic> infection. Hence we effectively installed a photo-activatable pyridinium warhead on the 5\u2032-OH of triazole-linked ribosylated ubiquitin allowing crosslinking of the probe to the catalytic histidine residues in <jats:italic>Legionella<\/jats:italic> SidE or Dup enzymes. <jats:italic>In vitro<\/jats:italic> tests on recombinantly expressed DupA and SdeA<jats:sub>PDE<\/jats:sub> revealed that the probe was able to capture the enzymes covalently upon photo-activation.<\/jats:p>","DOI":"10.3389\/fmolb.2024.1422034","type":"journal-article","created":{"date-parts":[[2024,7,9]],"date-time":"2024-07-09T04:47:07Z","timestamp":1720500427000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Capturing Legionella pneumophila effector enzymes using a ubiquitin derived photo-activatable probe"],"prefix":"10.3389","volume":"11","author":[{"given":"Max S.","family":"Kloet","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gerbrand J.","family":"van der Heden van Noort","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2024,7,9]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"6174","DOI":"10.1038\/s41467-021-26429-y","article-title":"Structural basis for protein glutamylation by the Legionella pseudokinase SidJ","volume":"12","author":"Adams","year":"2021","journal-title":"Nat. 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