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Novel assay for efficient iron release from ferritin

Ferritin plays an important role in iron detoxification and iron storage by converting soluble iron(II) to insoluble iron(III) inside the protein cage. Mechanism of in vitro ferritin iron release occurs more readily under reducing conditions, whereby reducing agents may enter into the ferritin cage...

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Main Authors:	Lim W.F., Abdullah M., Yap B.K., George E., Lai M.I.
Other Authors:	50061393300
Format:	Conference Paper
Published:	Institute of Electrical and Electronics Engineers Inc. 2023
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spelling	my.uniten.dspace-219922023-05-16T10:46:31Z Novel assay for efficient iron release from ferritin Lim W.F. Abdullah M. Yap B.K. George E. Lai M.I. 50061393300 55666914500 26649255900 7202827990 13605357700 Ferritin plays an important role in iron detoxification and iron storage by converting soluble iron(II) to insoluble iron(III) inside the protein cage. Mechanism of in vitro ferritin iron release occurs more readily under reducing conditions, whereby reducing agents may enter into the ferritin cage through the protein pores to convert insoluble iron(III) into soluble iron(II), after accepting an electron from reducing agents. The soluble iron(II) then liberates outside the protein cage and forms coloured and stable complexes in the presence of chromophore, an iron(II) chelating agent, to be quantified by spectrophotometry. However, iron(II) is readily converted back to iron(III) in the presence of oxygen. Thus, oxygen scavenger compounds were used to remove any dissolved oxygen. Studies suggested that low concentration of chaotropic agents regulate the protein pore opening and result in protein unfolding that is therefore accessible by reducing agents more effectively. According to in vivo studies, iron exit from ferritin for recycling or meeting the cell's need via lysosomal and/or proteasomal pathways. We designed an assay to determine the total iron available inside the ferritin by first using the commercially available iron(II) or iron(III) compounds as a substrate to optimize the concentration of reactants involved. Apart from chromophore and reducing agents, oxygen scavenger compounds, chaotropic agents and enzymes were added in order to induce ferritin iron release effectively. Linear calibration curve of the iron(II)-chromophore complex were obtained and was used as a reference to correlate absorbance readings to concentration of iron level. Reducing agents was able to reduce the iron(III) to iron(II) and oxygen scavenger A seems to be more effective in removing any dissolved oxygen to avoid oxidation of both the reducing agents and iron(III). Taken altogether, this experiment setup will be beneficial for the improvement of efficiency in quantifying the total iron level inside the ferritin. © 2014 IEEE. Final 2023-05-16T02:46:30Z 2023-05-16T02:46:30Z 2014 Conference Paper 10.1109/IECBES.2014.7047471 2-s2.0-84925582931 https://www.scopus.com/inward/record.uri?eid=2-s2.0-84925582931&doi=10.1109%2fIECBES.2014.7047471&partnerID=40&md5=441dae4955bd7bc3cce954abf537c157 https://irepository.uniten.edu.my/handle/123456789/21992 7047471 132 136 All Open Access, Green Institute of Electrical and Electronics Engineers Inc. Scopus
institution	Universiti Tenaga Nasional
building	UNITEN Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Tenaga Nasional
content_source	UNITEN Institutional Repository
url_provider	http://dspace.uniten.edu.my/
description	Ferritin plays an important role in iron detoxification and iron storage by converting soluble iron(II) to insoluble iron(III) inside the protein cage. Mechanism of in vitro ferritin iron release occurs more readily under reducing conditions, whereby reducing agents may enter into the ferritin cage through the protein pores to convert insoluble iron(III) into soluble iron(II), after accepting an electron from reducing agents. The soluble iron(II) then liberates outside the protein cage and forms coloured and stable complexes in the presence of chromophore, an iron(II) chelating agent, to be quantified by spectrophotometry. However, iron(II) is readily converted back to iron(III) in the presence of oxygen. Thus, oxygen scavenger compounds were used to remove any dissolved oxygen. Studies suggested that low concentration of chaotropic agents regulate the protein pore opening and result in protein unfolding that is therefore accessible by reducing agents more effectively. According to in vivo studies, iron exit from ferritin for recycling or meeting the cell's need via lysosomal and/or proteasomal pathways. We designed an assay to determine the total iron available inside the ferritin by first using the commercially available iron(II) or iron(III) compounds as a substrate to optimize the concentration of reactants involved. Apart from chromophore and reducing agents, oxygen scavenger compounds, chaotropic agents and enzymes were added in order to induce ferritin iron release effectively. Linear calibration curve of the iron(II)-chromophore complex were obtained and was used as a reference to correlate absorbance readings to concentration of iron level. Reducing agents was able to reduce the iron(III) to iron(II) and oxygen scavenger A seems to be more effective in removing any dissolved oxygen to avoid oxidation of both the reducing agents and iron(III). Taken altogether, this experiment setup will be beneficial for the improvement of efficiency in quantifying the total iron level inside the ferritin. © 2014 IEEE.
author2	50061393300
author_facet	50061393300 Lim W.F. Abdullah M. Yap B.K. George E. Lai M.I.
format	Conference Paper
author	Lim W.F. Abdullah M. Yap B.K. George E. Lai M.I.
spellingShingle	Lim W.F. Abdullah M. Yap B.K. George E. Lai M.I. Novel assay for efficient iron release from ferritin
author_sort	Lim W.F.
title	Novel assay for efficient iron release from ferritin
title_short	Novel assay for efficient iron release from ferritin
title_full	Novel assay for efficient iron release from ferritin
title_fullStr	Novel assay for efficient iron release from ferritin
title_full_unstemmed	Novel assay for efficient iron release from ferritin
title_sort	novel assay for efficient iron release from ferritin
publisher	Institute of Electrical and Electronics Engineers Inc.
publishDate	2023
_version_	1806428285846945792
score	13.211869

Novel assay for efficient iron release from ferritin

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