{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T06:53:18Z","timestamp":1790232798520,"version":"4.1.0"},"reference-count":43,"publisher":"National Academy of Sciences","issue":"12","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2009,3,24]]},"abstract":"<jats:p>\n                    Nitrosothiols (RSNO), formed from thiols and metabolites of nitric oxide (\u2022NO), have been implicated in a diverse set of physiological and pathophysiological processes, although the exact mechanisms by which they are formed biologically are unknown. Several candidate nitrosative pathways involve the reaction of \u2022NO with O\n                    <jats:sub>2<\/jats:sub>\n                    , reactive oxygen species (ROS), and transition metals. We developed a strategy using extracellular ferrocyanide to determine that under our conditions intracellular protein RSNO formation occurs from reaction of \u2022NO inside the cell, as opposed to cellular entry of nitrosative reactants from the extracellular compartment. Using this method we found that in RAW 264.7 cells RSNO formation occurs only at very low (&lt;8 \u03bcM) O\n                    <jats:sub>2<\/jats:sub>\n                    concentrations and exhibits zero-order dependence on \u2022NO concentration. Indeed, RSNO formation is not inhibited even at O\n                    <jats:sub>2<\/jats:sub>\n                    levels &lt;1 \u03bcM. Additionally, chelation of intracellular chelatable iron pool (CIP) reduces RSNO formation by &gt;50%. One possible metal-dependent, O\n                    <jats:sub>2<\/jats:sub>\n                    -independent nitrosative pathway is the reaction of thiols with dinitrosyliron complexes (DNIC), which are formed in cells from the reaction of \u2022NO with the CIP. Under our conditions, DNIC formation, like RSNO formation, is inhibited by \u224850% after chelation of labile iron. Both DNIC and RSNO are also increased during overproduction of ROS by the redox cycler 5,8-dimethoxy-1,4-naphthoquinone. Taken together, these data strongly suggest that cellular RSNO are formed from free \u2022NO via transnitrosation from DNIC derived from the CIP. We have examined in detail the kinetics and mechanism of RSNO formation inside cells.\n                  <\/jats:p>","DOI":"10.1073\/pnas.0710416106","type":"journal-article","created":{"date-parts":[[2009,3,4]],"date-time":"2009-03-04T21:40:29Z","timestamp":1236202829000},"page":"4671-4676","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":197,"title":["Dinitrosyliron complexes and the mechanism(s) of cellular protein nitrosothiol formation from nitric oxide"],"prefix":"10.1073","volume":"106","author":[{"given":"Charles A.","family":"Bosworth","sequence":"first","affiliation":[{"name":"Departments of aPhysiology and Biophysics,"},{"name":"Center for Free Radical Biology, University of Alabama, Birmingham, AL 35205; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"suffix":"Jr.","given":"Jos\u00e9 C.","family":"Toledo","sequence":"additional","affiliation":[{"name":"Centro de Ci\u00eancias Naturais e Humanas, Universidade Federal do ABC, CEP 09210-170, Santo Andr\u00e9, Sao Paulo, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jaroslaw W.","family":"Zmijewski","sequence":"additional","affiliation":[{"name":"Medicine,"},{"name":"Center for Free Radical Biology, University of Alabama, Birmingham, AL 35205; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qian","family":"Li","sequence":"additional","affiliation":[{"name":"Anesthesiology, and"},{"name":"Center for Free Radical Biology, University of Alabama, Birmingham, AL 35205; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"suffix":"Jr.","given":"Jack R.","family":"Lancaster","sequence":"additional","affiliation":[{"name":"Departments of aPhysiology and Biophysics,"},{"name":"Anesthesiology, and"},{"name":"Environmental Health Sciences, and"},{"name":"Center for Free Radical Biology, University of Alabama, Birmingham, AL 35205; and"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2009,3,24]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0306706101"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrm1569"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.freeradbiomed.2004.12.016"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.270.47.28158"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1080\/15216540701311147"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.5.2841"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.152157599"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.270.49.29244"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1006\/niox.1997.0122"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.274.38.27069"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.275.15.10954"},{"key":"e_1_3_3_12_2","first-page":"313","article-title":"Effects of metal ions on the oxidation and nitrosation of cysteine residues in proteins and enzymes","volume":"38","author":"English AM","year":"2001","unstructured":"AM English, DE Wilcox, Effects of metal ions on the oxidation and nitrosation of cysteine residues in proteins and enzymes. 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