{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T17:22:14Z","timestamp":1791566534241,"version":"4.3.4"},"reference-count":54,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2015,2,24]],"date-time":"2015-02-24T00:00:00Z","timestamp":1424736000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100003339","name":"Consejo Superior de Investigaciones Cient\u00edficas","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003339","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Plant Journal"],"published-print":{"date-parts":[[2015,3]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>Protein tyrosine (Tyr) nitration is a post\u2010translational modification yielding 3\u2010nitrotyrosine (NO<jats:sub>2<\/jats:sub>\u2013Tyr). Formation of <jats:styled-content style=\"fixed-case\">NO<\/jats:styled-content><jats:sub>2<\/jats:sub>\u2013Tyr is generally considered as a marker of nitro\u2010oxidative stress and is involved in some human pathophysiological disorders, but has been poorly studied in plants. Leghemoglobin (Lb) is an abundant hemeprotein of legume nodules that plays an essential role as an O<jats:sub>2<\/jats:sub> transporter. Liquid chromatography coupled to tandem mass spectrometry was used for a targeted search and quantification of <jats:styled-content style=\"fixed-case\">NO<\/jats:styled-content><jats:sub>2<\/jats:sub>\u2013Tyr in Lb. For all Lbs examined, Tyr30, located in the distal heme pocket, is the major target of nitration. Lower amounts were found for <jats:styled-content style=\"fixed-case\">NO<\/jats:styled-content><jats:sub>2<\/jats:sub>\u2013Tyr25 and <jats:styled-content style=\"fixed-case\">NO<\/jats:styled-content><jats:sub>2<\/jats:sub>\u2013Tyr133. Nitrated Lb and other as yet unidentified nitrated proteins were also detected in nodules of plants not receiving <jats:inline-graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"graphic\/tpj12762-math-0001.png\" xlink:title=\"urn:x-wiley:09607412:media:tpj12762:tpj12762-math-0001\"\/> and were found to decrease during senescence. This demonstrates formation of nitric oxide (\u02d9<jats:styled-content style=\"fixed-case\">NO<\/jats:styled-content>) and <jats:inline-graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"graphic\/tpj12762-math-0002.png\" xlink:title=\"urn:x-wiley:09607412:media:tpj12762:tpj12762-math-0002\"\/> by alternative means to nitrate reductase, probably via a \u02d9<jats:styled-content style=\"fixed-case\">NO<\/jats:styled-content> synthase\u2010like enzyme, and strongly suggests that nitrated proteins perform biological functions and are not merely metabolic byproducts. <jats:italic>In vitro<\/jats:italic> assays with purified Lb revealed that Tyr nitration requires <jats:inline-graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"graphic\/tpj12762-math-0003.png\" xlink:title=\"urn:x-wiley:09607412:media:tpj12762:tpj12762-math-0003\"\/>\u00a0+\u00a0H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> and that peroxynitrite is not an efficient inducer of nitration, probably because Lb isomerizes it to <jats:inline-graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"graphic\/tpj12762-math-0004.png\" xlink:title=\"urn:x-wiley:09607412:media:tpj12762:tpj12762-math-0004\"\/>. Nitrated Lb is formed via oxoferryl Lb, which generates nitrogen dioxide and tyrosyl radicals. This mechanism is distinctly different from that involved in heme nitration. Formation of <jats:styled-content style=\"fixed-case\">NO<\/jats:styled-content><jats:sub>2<\/jats:sub>\u2013Tyr in Lb is a consequence of active metabolism in functional nodules, where Lb may act as a sink of toxic peroxynitrite and may play a protective role in the symbiosis.<\/jats:p>","DOI":"10.1111\/tpj.12762","type":"journal-article","created":{"date-parts":[[2015,1,20]],"date-time":"2015-01-20T13:50:58Z","timestamp":1421761858000},"page":"723-735","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":65,"title":["Leghemoglobin is nitrated in functional legume nodules in a tyrosine residue within the heme cavity by a nitrite\/peroxide\u2010dependent mechanism"],"prefix":"10.1111","volume":"81","author":[{"given":"Martha","family":"Sainz","sequence":"first","affiliation":[{"name":"Departamento de Nutrici\u00f3n Vegetal, Estaci\u00f3n Experimental de Aula Dei Consejo Superior de Investigaciones Cient\u00edficas (CSIC) 50080 Zaragoza Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura","family":"Calvo\u2010Begueria","sequence":"additional","affiliation":[{"name":"Departamento de Nutrici\u00f3n Vegetal, Estaci\u00f3n Experimental de Aula Dei Consejo Superior de Investigaciones Cient\u00edficas (CSIC) 50080 Zaragoza Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carmen","family":"P\u00e9rez\u2010Rontom\u00e9","sequence":"additional","affiliation":[{"name":"Departamento de Nutrici\u00f3n Vegetal, Estaci\u00f3n Experimental de Aula Dei Consejo Superior de Investigaciones Cient\u00edficas (CSIC) 50080 Zaragoza Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefanie","family":"Wienkoop","sequence":"additional","affiliation":[{"name":"Department of Ecogenomics and Systems Biology University of Vienna 1090 Vienna Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joaqu\u00edn","family":"Abi\u00e1n","sequence":"additional","affiliation":[{"name":"Laboratorio de Prote\u00f3mica CSIC\u2010Universidad Aut\u00f3noma de Barcelona Instituto de Investigaciones Biom\u00e9dicas de Barcelona 08036 Barcelona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christiana","family":"Staudinger","sequence":"additional","affiliation":[{"name":"Laboratorio de Prote\u00f3mica CSIC\u2010Universidad Aut\u00f3noma de Barcelona Instituto de Investigaciones Biom\u00e9dicas de Barcelona 08036 Barcelona Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvina","family":"Bartesaghi","sequence":"additional","affiliation":[{"name":"Departamento de Bioqu\u00edmica and Center for Free Radical and Biomedical Research Facultad de Medicina Universidad de la Rep\u00fablica Montevideo 11800 Uruguay"},{"name":"Departamento de Educaci\u00f3n M\u00e9dica Facultad de Medicina Universidad de la Rep\u00fablica Montevideo 11800 Uruguay"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rafael","family":"Radi","sequence":"additional","affiliation":[{"name":"Departamento de Bioqu\u00edmica and Center for Free Radical and Biomedical Research Facultad de Medicina Universidad de la Rep\u00fablica Montevideo 11800 Uruguay"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manuel","family":"Becana","sequence":"additional","affiliation":[{"name":"Departamento de Nutrici\u00f3n Vegetal, Estaci\u00f3n Experimental de Aula Dei Consejo Superior de Investigaciones Cient\u00edficas (CSIC) 50080 Zaragoza Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2015,2,24]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(02)59198-9"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.147678"},{"key":"e_1_2_6_4_1","first-page":"315","volume-title":"Methods for Evaluating Biological Nitrogen Fixation","author":"Appleby C.A.","year":"1980"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1093\/jxb\/49.318.41"},{"key":"e_1_2_6_6_1","doi-asserted-by":"crossref","first-page":"5685","DOI":"10.1016\/S0021-9258(17)30321-6","article-title":"The reaction of ferrous leghemoglobin with hydrogen peroxide to form leghemoglobin (IV)","volume":"253","author":"Aviram I.","year":"1978","journal-title":"J. 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