{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T12:53:02Z","timestamp":1791895982740,"version":"4.3.4"},"reference-count":52,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T00:00:00Z","timestamp":1786492800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T00:00:00Z","timestamp":1786492800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001411","name":"Indian Council of Medical Research","doi-asserted-by":"publisher","award":["Ref. EM\/Dev\/SG\/75\/0782\/2023"],"award-info":[{"award-number":["Ref. 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Hemoglobin (Hb), the principal oxygen\u2010transport protein of erythrocytes, is a key intravascular scavenger and target of PN; however, how controlled PN exposure translates into structural remodeling and aggregation of Hb remains poorly understood. Purified human Hb was exposed to PN across sub\u2010stoichiometric to moderate oxidant excess and characterized using intrinsic fluorescence, UV\u2013visible (UV\u2013vis) spectroscopy, 2,4\u2010dinitrophenylhydrazine (DNPH) carbonyl assay, Thioflavin T (ThT), 8\u2010anilino\u20101\u2010naphthalenesulfonic acid (ANS), Congo Red (CR), Fourier\u2010transform infrared (FTIR) spectroscopy with Amide I deconvolution, dynamic light scattering (DLS) with zeta potential, x\u2010ray powder diffraction (XRPD), scanning electron microscopy (SEM), atomic force microscopy (AFM), and fluorescence microscopy. At low oxidant loads, PN caused concentration\u2010dependent fluorescence quenching, carbonyl accumulation, and graded UV\u2013visible absorbance increases, consistent with heme\u2010centered oxidation and limited aromatic residue modification. At higher concentrations, Amide I deconvolution revealed collapse of\n                    <jats:italic>\u03b1<\/jats:italic>\n                    \u2010helical content from 66.41% to 8.81% and\n                    <jats:italic>\u03b2<\/jats:italic>\n                    \u2010sheet enrichment to 71.18%, accompanied by surface charge neutralization, increased hydrodynamic diameter, and enhanced nanoscale roughness. Despite\n                    <jats:italic>\u03b2<\/jats:italic>\n                    \u2010sheet accumulation and ThT\/CR\u2010positive aggregate formation, XRPD, AFM, SEM, and fluorescence microscopy showed no long\u2010range crystalline order or fibrillar morphology, demonstrating that PN drives Hb toward amorphous and oligomeric assemblies rather than canonical amyloid fibrils. Time\u2010dependent assays at 37\u00b0C confirmed that PN modification lowers the nucleation barrier for thermally driven aggregation. These findings define physicochemical thresholds separating protective scavenging from structural destabilization and offer a framework for distinguishing nitroxidative protein aggregation from classical amyloid fibrillation.\n                  <\/jats:p>","DOI":"10.1002\/pro.70762","type":"journal-article","created":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T10:35:06Z","timestamp":1786530906000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Peroxynitrite\u2010induced structural remodeling and aggregation of hemoglobin under nitroxidative stress"],"prefix":"10.1002","volume":"35","author":[{"given":"Shaik","family":"Basha","sequence":"first","affiliation":[{"name":"Department of Biophysics Manipal School of Life Sciences, Manipal Academy of Higher Education  Manipal 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