H ferritin silencing induces protein misfolding in K562 cells: A Raman analysis

Handle URI:
http://hdl.handle.net/10754/581665
Title:
H ferritin silencing induces protein misfolding in K562 cells: A Raman analysis
Authors:
Zolea, Fabiana; Biamonte, Flavia; Candeloro, Patrizio; Di Sanzo, Maddalena; Cozzi, Anna; Di Vito, Anna; Quaresima, Barbara; Lobello, Nadia; Trecroci, Francesca; Di Fabrizio, Enzo M. ( 0000-0001-5886-4678 ) ; Levi, Sonia; Cuda, Giovanni ( 0000-0001-6313-1866 ) ; Costanzo, Francesco
Abstract:
The redox state of the cell is involved in the regulation of many physiological functions as well as in the pathogenesis of several diseases, and is strictly dependent on the amount of iron in its catalytically active state. Alterations of iron homeostasis determine increased steady-state concentrations of Reactive Oxygen Species (ROS) that cause lipid peroxidation, DNA damage and altered protein folding. Ferritin keeps the intracellular iron in a non-toxic and readily available form and consequently plays a central role in iron and redox homeostasis. The protein is composed by 24 subunits of the H- and L-type, coded by two different genes, with structural and functional differences. The aim of this study was to shed light on the role of the single H ferritin subunit (FHC) in keeping the native correct protein three-dimensional structure. To this, we performed Raman spectroscopy on protein extracts from K562 cells subjected to FHC silencing. The results show a significant increase in the percentage of disordered structures content at a level comparable to that induced by H2O2 treatment in control cells. ROS inhibitor and iron chelator were able to revert protein misfolding. This integrated approach, involving Raman spectroscopy and targeted-gene silencing, indicates that an imbalance of the heavy-to-light chain ratio in the ferritin composition is able to induce severe but still reversible modifications in protein folding and uncovers new potential pathogenetic mechanisms associated to intracellular iron perturbation.
KAUST Department:
Physical Sciences and Engineering (PSE) Division
Citation:
H ferritin silencing induces protein misfolding in K562 cells: A Raman analysis 2015 Free Radical Biology and Medicine
Publisher:
Elsevier BV
Journal:
Free Radical Biology and Medicine
Issue Date:
9-Oct-2015
DOI:
10.1016/j.freeradbiomed.2015.07.161
Type:
Article
ISSN:
08915849
Additional Links:
http://linkinghub.elsevier.com/retrieve/pii/S0891584915005900
Appears in Collections:
Articles; Physical Sciences and Engineering (PSE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorZolea, Fabianaen
dc.contributor.authorBiamonte, Flaviaen
dc.contributor.authorCandeloro, Patrizioen
dc.contributor.authorDi Sanzo, Maddalenaen
dc.contributor.authorCozzi, Annaen
dc.contributor.authorDi Vito, Annaen
dc.contributor.authorQuaresima, Barbaraen
dc.contributor.authorLobello, Nadiaen
dc.contributor.authorTrecroci, Francescaen
dc.contributor.authorDi Fabrizio, Enzo M.en
dc.contributor.authorLevi, Soniaen
dc.contributor.authorCuda, Giovannien
dc.contributor.authorCostanzo, Francescoen
dc.date.accessioned2015-11-04T06:41:01Zen
dc.date.available2015-11-04T06:41:01Zen
dc.date.issued2015-10-09en
dc.identifier.citationH ferritin silencing induces protein misfolding in K562 cells: A Raman analysis 2015 Free Radical Biology and Medicineen
dc.identifier.issn08915849en
dc.identifier.doi10.1016/j.freeradbiomed.2015.07.161en
dc.identifier.urihttp://hdl.handle.net/10754/581665en
dc.description.abstractThe redox state of the cell is involved in the regulation of many physiological functions as well as in the pathogenesis of several diseases, and is strictly dependent on the amount of iron in its catalytically active state. Alterations of iron homeostasis determine increased steady-state concentrations of Reactive Oxygen Species (ROS) that cause lipid peroxidation, DNA damage and altered protein folding. Ferritin keeps the intracellular iron in a non-toxic and readily available form and consequently plays a central role in iron and redox homeostasis. The protein is composed by 24 subunits of the H- and L-type, coded by two different genes, with structural and functional differences. The aim of this study was to shed light on the role of the single H ferritin subunit (FHC) in keeping the native correct protein three-dimensional structure. To this, we performed Raman spectroscopy on protein extracts from K562 cells subjected to FHC silencing. The results show a significant increase in the percentage of disordered structures content at a level comparable to that induced by H2O2 treatment in control cells. ROS inhibitor and iron chelator were able to revert protein misfolding. This integrated approach, involving Raman spectroscopy and targeted-gene silencing, indicates that an imbalance of the heavy-to-light chain ratio in the ferritin composition is able to induce severe but still reversible modifications in protein folding and uncovers new potential pathogenetic mechanisms associated to intracellular iron perturbation.en
dc.language.isoenen
dc.publisherElsevier BVen
dc.relation.urlhttp://linkinghub.elsevier.com/retrieve/pii/S0891584915005900en
dc.rightsNOTICE: this is the author’s version of a work that was accepted for publication in Free Radical Biology and Medicine. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Free Radical Biology and Medicine, 9 October 2015. DOI: 10.1016/j.freeradbiomed.2015.07.161en
dc.subjectFerritin heavy chainen
dc.subjectOxidative stressen
dc.subjectProtein misfoldingen
dc.subjectRaman spectroscopyen
dc.subjectK562en
dc.titleH ferritin silencing induces protein misfolding in K562 cells: A Raman analysisen
dc.typeArticleen
dc.contributor.departmentPhysical Sciences and Engineering (PSE) Divisionen
dc.identifier.journalFree Radical Biology and Medicineen
dc.eprint.versionPost-printen
dc.contributor.institutionDepartment of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Catanzaro, Italy, 88100en
dc.contributor.institutionBioNEM Laboratory, Department of Experimental and Clinical Medicine, Magna Græcia University of Catanzaro, Salvatore Venuta Campus, Catanzaro, Italy, 88100en
dc.contributor.institutionSan Raffaele Scientific Institute, Division of Neuroscience, Milano, Italy, 20132en
dc.contributor.institutionUniversity Vita-Salute San Raffaele, Milano, Italy, 20132en
dc.contributor.affiliationKing Abdullah University of Science and Technology (KAUST)en
kaust.authorDi Fabrizio, Enzo M.en
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