{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T00:37:49Z","timestamp":1783039069070,"version":"3.54.6"},"reference-count":39,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2024,4,9]],"date-time":"2024-04-09T00:00:00Z","timestamp":1712620800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2024,4,9]],"date-time":"2024-04-09T00:00:00Z","timestamp":1712620800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2024,5]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Chemical warfare nerve agents and pesticides, known as organophosphorus compounds inactivate cholinesterases (ChEs) by phosphorylating the serine hydroxyl group located at the active site of ChEs. Over the course of time, phosphorylation is followed by loss of an organophosphate\u2010leaving group and the bond with ChEs becomes irreversible, a process known as aging. Differently, structurally related irreversible catalytic poisons bearing sulfur instead of phosphorus convert ChEs in its aged form only by covalently binding to the key catalytic serine. Kinetic and crystallographic studies of the interaction between\n                    <jats:italic>Torpedo californica<\/jats:italic>\n                    acetylcholinesterase (\n                    <jats:italic>Tc<\/jats:italic>\n                    AChE) and a small organosulfonate, methanesulfonyl fluoride (MSF), indeed revealed irreversibly methylsulfonylated serine 200, to be isosteric with the bound aged sarin\/soman analogues. The potent bulky reversible inhibitor 7\u2010bis\u2010tacrine (BTA) adopts, in the active site of the crystal structure of the MSF\u2010enzyme adduct, a location and an orientation that closely resemble the one being found in the crystal structure of the BTA\u2010enzyme complex. Remarkably, the presence of BTA accelerates the rate of methanesulfonylation by a factor of two. This unexpected result can be explained on the basis of two facts: i) the steric hindrance exerted by BTA to MSF in accessing the active site and ii) the acceleration of the MSF\u2010enzyme adduct formation as a consequence of the lowering of the rotational and translational degrees of freedom in the proximity of the catalytic serine. It is well known that pralidoxime (2\u2010Pyridine Aldoxime Methyl chloride, 2\u2010PAM) alone or in the presence of the substrate acetylcholine cannot reactivate the active site serine of the\n                    <jats:italic>Tc<\/jats:italic>\n                    AChE\u2010MSF adduct. We show that the simultaneous presence of 2\u2010PAM and the additional neutral oxime, 2\u2010[(hydroxyimino)methyl]\u2010l\u2010methylimidazol (2\u2010HAM), triggers the reactivation process of\n                    <jats:italic>Tc<\/jats:italic>\n                    AChE within the hour timescale. Overall, our results pave the way toward the likely use of a cocktail of distinctive oximes as a promising recipe for an effective and fast reactivation of aged cholinesterases.\n                  <\/jats:p>","DOI":"10.1002\/pro.4977","type":"journal-article","created":{"date-parts":[[2024,4,9]],"date-time":"2024-04-09T06:55:14Z","timestamp":1712645714000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Disentangling the formation, mechanism, and evolvement of the covalent methanesulfonyl fluoride acetylcholinesterase adduct: Insights into an aged\u2010like inactive complex susceptible to reactivation by a combination of nucleophiles"],"prefix":"10.1002","volume":"33","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6940-0550","authenticated-orcid":false,"given":"Jure","family":"Stojan","sequence":"first","affiliation":[{"name":"Institute of Biochemistry, Faculty of Medicine University of Ljubljana  Ljubljana Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9662-064X","authenticated-orcid":false,"given":"Alessandro","family":"Pesaresi","sequence":"additional","affiliation":[{"name":"Trieste Outstation, Area Science Park\u2010Basovizza Institute of Crystallography, National Research Council  Trieste Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"An\u017ee","family":"Meden","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy University of Ljubljana  Ljubljana Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6859-7868","authenticated-orcid":false,"given":"Doriano","family":"Lamba","sequence":"additional","affiliation":[{"name":"Trieste Outstation, Area Science Park\u2010Basovizza Institute of Crystallography, National Research Council  Trieste Italy"},{"name":"Interuniversity Consortium \u201cBiostructures and Biosystems National 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