{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T07:49:04Z","timestamp":1791964144378,"version":"4.3.4"},"reference-count":41,"publisher":"Wiley","issue":"19","license":[{"start":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T00:00:00Z","timestamp":1790640000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T00:00:00Z","timestamp":1790640000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100005283","name":"FUNCAP","doi-asserted-by":"publisher","award":["PS1\u20100186\u201000087.01.00\/21"],"award-info":[{"award-number":["PS1\u20100186\u201000087.01.00\/21"]}],"id":[{"id":"10.13039\/501100005283","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemBioChem"],"published-print":{"date-parts":[[2026,10,14]]},"abstract":"<jats:p>Nitric oxide (NO) and nitroxyl (HNO) are closely related reactive nitrogen species with distinct biological functions, yet differentiating their chemistry remains challenging because of their high reactivity and overlapping reaction pathways. Developing simple protein\u2010based platforms to study their reactivity is therefore of considerable interest. Rubredoxins are small, stable iron\u2013thiolate proteins widely used as model systems for investigating metal\u2013thiolate coordination and electron transfer. Here, we investigated the reactivity of native iron\u2010rubredoxin (Fe\u2010Rd) and its nickel\u2010substituted analog (Ni\u2010Rd) toward NO and HNO to determine whether cysteine ligand modification could disrupt metal\u2013thiolate coordination and induce demetallation. Electronic absorption and fluorescence spectroscopy showed that Fe\u2010Rd is highly resistant to both species, whereas Ni\u2010Rd exhibited pronounced spectral changes consistent with metal release, including loss of the Ni\u2500S ligand\u2010to\u2010metal charge\u2010transfer band and increased intrinsic fluorescence. Inductively coupled plasma atomic emission spectroscopy confirmed partial nickel release following treatment with both NO and HNO donors. Despite demetallation, circular dichroism, thermal denaturation, and size\u2010exclusion chromatography demonstrated that Ni\u2010Rd largely retains its secondary structure, thermal stability, and monomeric state. These findings establish Ni\u2010Rd as a robust minimal metalloprotein platform for probing reactive nitrogen species\u2010induced thiolate chemistry.<\/jats:p>","DOI":"10.1002\/cbic.70548","type":"journal-article","created":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T17:27:04Z","timestamp":1790702824000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Nickel\u2010Substituted Rubredoxin Undergoes Demetallation Upon Reaction With Nitric Oxide and Nitroxyl"],"prefix":"10.1002","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0007-8452","authenticated-orcid":false,"given":"Eduardo H. 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M.","family":"da Silva","sequence":"additional","affiliation":[{"name":"Bioinorganic Group Department of Organic and Inorganic Chemistry Federal University of Cear\u00e1  Fortaleza Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3222-8141","authenticated-orcid":false,"given":"Marta S. P.","family":"Carepo","sequence":"additional","affiliation":[{"name":"LAQV REQUIMTE Departamento de Qui\u0301mica Faculdade de Cie\u0302ncias e Tecnologia Universidade Nova de Lisboa  Caparica Portugal"},{"name":"Escola de Psicologia e Ci\u00eancias da Vida Departamento de Ci\u00eancias da Vida Universidade Lus\u00f3fona  Lisboa Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2149-9416","authenticated-orcid":false,"given":"Sofia R.","family":"Pauleta","sequence":"additional","affiliation":[{"name":"Microbial Stress Lab UCIBIO Departamento de Qui\u0301mica Faculdade de Cie\u0302ncias e Tecnologia Universidade Nova de Lisboa  Caparica Portugal"},{"name":"i4HB Departamento de Qui\u0301mica Faculdade de Cie\u0302ncias e Tecnologia Universidade Nova de Lisboa  Caparica Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0971-4977","authenticated-orcid":false,"given":"Isabel","family":"Moura","sequence":"additional","affiliation":[{"name":"LAQV REQUIMTE Departamento de Qui\u0301mica Faculdade de Cie\u0302ncias e Tecnologia Universidade Nova de Lisboa  Caparica Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4726-2388","authenticated-orcid":false,"given":"Jos\u00e9 J. 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