Effect of macromolecular crowding on protein oxidation: Consequences on the rate, extent and oxidation pathways

Biological systems are heterogeneous and crowded environments. Such packed milieus are expected to modulate reactions both inside and outside the cell, including protein oxidation. In this work, we explored the effect of macromolecular crowding on the rate and extent of oxidation of Trp and Tyr, in...

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Autores principales:	Eduardo Fuentes-Lemus, Juan Sebastián Reyes, Luke F. Gamon, Camilo López-Alarcón, Michael J. Davies
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Macromolecular crowding Protein oxidation Chain reactions Tryptophan Tyrosine Peroxyl radicals Medicine (General) R5-920 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/d126413da117403fb505baa65ea369bf
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spelling	oai:doaj.org-article:d126413da117403fb505baa65ea369bf2021-12-02T05:01:31ZEffect of macromolecular crowding on protein oxidation: Consequences on the rate, extent and oxidation pathways2213-231710.1016/j.redox.2021.102202https://doaj.org/article/d126413da117403fb505baa65ea369bf2021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2213231721003621https://doaj.org/toc/2213-2317Biological systems are heterogeneous and crowded environments. Such packed milieus are expected to modulate reactions both inside and outside the cell, including protein oxidation. In this work, we explored the effect of macromolecular crowding on the rate and extent of oxidation of Trp and Tyr, in free amino acids, peptides and proteins. These species were chosen as they are readily oxidized and contribute to damage propagation. Dextran was employed as an inert crowding agent, as this polymer decreases the fraction of volume available to other (macro)molecules. Kinetic analysis demonstrated that dextran enhanced the rate of oxidation of free Trp, and peptide Trp, elicited by AAPH-derived peroxyl radicals. For free Trp, the rates of oxidation were 15.0 ± 2.1 and 30.5 ± 3.4 μM min−1 without and with dextran (60 mg mL−1) respectively. Significant increases were also detected for peptide-incorporated Trp. Dextran increased the extent of Trp consumption (up to 2-fold) and induced short chain reactions. In contrast, Tyr oxidation was not affected by the presence of dextran. Studies on proteins, using SDS-PAGE and LC-MS, indicated that oxidation was also affected by crowding, with enhanced amino acid loss (45% for casein), chain reactions and altered extents of oligomer formation. The overall effects of dextran-mediated crowding were however dependent on the protein structure. Overall, these data indicate that molecular crowding, as commonly encountered in biological systems affect the rates, and extents of oxidation, and particularly of Trp residues, illustrating the importance of appropriate choice of in vitro systems to study biological oxidations.Eduardo Fuentes-LemusJuan Sebastián ReyesLuke F. GamonCamilo López-AlarcónMichael J. DaviesElsevierarticleMacromolecular crowdingProtein oxidationChain reactionsTryptophanTyrosinePeroxyl radicalsMedicine (General)R5-920Biology (General)QH301-705.5ENRedox Biology, Vol 48, Iss , Pp 102202- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Macromolecular crowding Protein oxidation Chain reactions Tryptophan Tyrosine Peroxyl radicals Medicine (General) R5-920 Biology (General) QH301-705.5
spellingShingle	Macromolecular crowding Protein oxidation Chain reactions Tryptophan Tyrosine Peroxyl radicals Medicine (General) R5-920 Biology (General) QH301-705.5 Eduardo Fuentes-Lemus Juan Sebastián Reyes Luke F. Gamon Camilo López-Alarcón Michael J. Davies Effect of macromolecular crowding on protein oxidation: Consequences on the rate, extent and oxidation pathways
description	Biological systems are heterogeneous and crowded environments. Such packed milieus are expected to modulate reactions both inside and outside the cell, including protein oxidation. In this work, we explored the effect of macromolecular crowding on the rate and extent of oxidation of Trp and Tyr, in free amino acids, peptides and proteins. These species were chosen as they are readily oxidized and contribute to damage propagation. Dextran was employed as an inert crowding agent, as this polymer decreases the fraction of volume available to other (macro)molecules. Kinetic analysis demonstrated that dextran enhanced the rate of oxidation of free Trp, and peptide Trp, elicited by AAPH-derived peroxyl radicals. For free Trp, the rates of oxidation were 15.0 ± 2.1 and 30.5 ± 3.4 μM min−1 without and with dextran (60 mg mL−1) respectively. Significant increases were also detected for peptide-incorporated Trp. Dextran increased the extent of Trp consumption (up to 2-fold) and induced short chain reactions. In contrast, Tyr oxidation was not affected by the presence of dextran. Studies on proteins, using SDS-PAGE and LC-MS, indicated that oxidation was also affected by crowding, with enhanced amino acid loss (45% for casein), chain reactions and altered extents of oligomer formation. The overall effects of dextran-mediated crowding were however dependent on the protein structure. Overall, these data indicate that molecular crowding, as commonly encountered in biological systems affect the rates, and extents of oxidation, and particularly of Trp residues, illustrating the importance of appropriate choice of in vitro systems to study biological oxidations.
format	article
author	Eduardo Fuentes-Lemus Juan Sebastián Reyes Luke F. Gamon Camilo López-Alarcón Michael J. Davies
author_facet	Eduardo Fuentes-Lemus Juan Sebastián Reyes Luke F. Gamon Camilo López-Alarcón Michael J. Davies
author_sort	Eduardo Fuentes-Lemus
title	Effect of macromolecular crowding on protein oxidation: Consequences on the rate, extent and oxidation pathways
title_short	Effect of macromolecular crowding on protein oxidation: Consequences on the rate, extent and oxidation pathways
title_full	Effect of macromolecular crowding on protein oxidation: Consequences on the rate, extent and oxidation pathways
title_fullStr	Effect of macromolecular crowding on protein oxidation: Consequences on the rate, extent and oxidation pathways
title_full_unstemmed	Effect of macromolecular crowding on protein oxidation: Consequences on the rate, extent and oxidation pathways
title_sort	effect of macromolecular crowding on protein oxidation: consequences on the rate, extent and oxidation pathways
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/d126413da117403fb505baa65ea369bf
work_keys_str_mv	AT eduardofuenteslemus effectofmacromolecularcrowdingonproteinoxidationconsequencesontherateextentandoxidationpathways AT juansebastianreyes effectofmacromolecularcrowdingonproteinoxidationconsequencesontherateextentandoxidationpathways AT lukefgamon effectofmacromolecularcrowdingonproteinoxidationconsequencesontherateextentandoxidationpathways AT camilolopezalarcon effectofmacromolecularcrowdingonproteinoxidationconsequencesontherateextentandoxidationpathways AT michaeljdavies effectofmacromolecularcrowdingonproteinoxidationconsequencesontherateextentandoxidationpathways
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Effect of macromolecular crowding on protein oxidation: Consequences on the rate, extent and oxidation pathways

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