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Substitution of histidine 30 by asparagine in manganese superoxide dismutase alters biophysical properties and supports proliferation in a K562 leukemia cell line

We have generated a mutant of C. elegans manganese superoxide dismutase at histidine 30 by site-directed mutagenesis. The structure was solved at a resolution of 1.52 Å by X-ray crystallography (pdb: 6S0D). His30 was targeted, as it forms as a gateway residue at the top of the solvent access funnel...

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Autores principales: Bonetta, Rosalin, Hunter, Gary J., Trinh, Chi H., Borowski, Tomasz, Fenech, Anthony G., Kulp, Maria, Tabares, Leandro C., Un, Sun, Hunter, Thérèse
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8190026/
https://www.ncbi.nlm.nih.gov/pubmed/34021366
http://dx.doi.org/10.1007/s00249-021-01544-2
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author Bonetta, Rosalin
Hunter, Gary J.
Trinh, Chi H.
Borowski, Tomasz
Fenech, Anthony G.
Kulp, Maria
Tabares, Leandro C.
Un, Sun
Hunter, Thérèse
author_facet Bonetta, Rosalin
Hunter, Gary J.
Trinh, Chi H.
Borowski, Tomasz
Fenech, Anthony G.
Kulp, Maria
Tabares, Leandro C.
Un, Sun
Hunter, Thérèse
author_sort Bonetta, Rosalin
collection PubMed
description We have generated a mutant of C. elegans manganese superoxide dismutase at histidine 30 by site-directed mutagenesis. The structure was solved at a resolution of 1.52 Å by X-ray crystallography (pdb: 6S0D). His30 was targeted, as it forms as a gateway residue at the top of the solvent access funnel to the active site, together with Tyr34. In the wild-type protein, these gateway residues are involved in the hydrogen-bonding network providing the protons necessary for the catalytic reaction at the metal center. However, biophysical characterization and cell viability experiments reveal that a mutation from histidine to asparagine in the H30N mutant modifies metal selectivity in the protein, favoring the uptake of iron over manganese in minimal media conditions, alters active-site coordination from the characteristic trigonal bipyramidal to octahedral geometry, and encourages cellular proliferation in K562 cells, when added exogenously to the cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00249-021-01544-2.
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spelling pubmed-81900262021-06-28 Substitution of histidine 30 by asparagine in manganese superoxide dismutase alters biophysical properties and supports proliferation in a K562 leukemia cell line Bonetta, Rosalin Hunter, Gary J. Trinh, Chi H. Borowski, Tomasz Fenech, Anthony G. Kulp, Maria Tabares, Leandro C. Un, Sun Hunter, Thérèse Eur Biophys J Original Article We have generated a mutant of C. elegans manganese superoxide dismutase at histidine 30 by site-directed mutagenesis. The structure was solved at a resolution of 1.52 Å by X-ray crystallography (pdb: 6S0D). His30 was targeted, as it forms as a gateway residue at the top of the solvent access funnel to the active site, together with Tyr34. In the wild-type protein, these gateway residues are involved in the hydrogen-bonding network providing the protons necessary for the catalytic reaction at the metal center. However, biophysical characterization and cell viability experiments reveal that a mutation from histidine to asparagine in the H30N mutant modifies metal selectivity in the protein, favoring the uptake of iron over manganese in minimal media conditions, alters active-site coordination from the characteristic trigonal bipyramidal to octahedral geometry, and encourages cellular proliferation in K562 cells, when added exogenously to the cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00249-021-01544-2. Springer International Publishing 2021-05-21 2021 /pmc/articles/PMC8190026/ /pubmed/34021366 http://dx.doi.org/10.1007/s00249-021-01544-2 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Original Article
Bonetta, Rosalin
Hunter, Gary J.
Trinh, Chi H.
Borowski, Tomasz
Fenech, Anthony G.
Kulp, Maria
Tabares, Leandro C.
Un, Sun
Hunter, Thérèse
Substitution of histidine 30 by asparagine in manganese superoxide dismutase alters biophysical properties and supports proliferation in a K562 leukemia cell line
title Substitution of histidine 30 by asparagine in manganese superoxide dismutase alters biophysical properties and supports proliferation in a K562 leukemia cell line
title_full Substitution of histidine 30 by asparagine in manganese superoxide dismutase alters biophysical properties and supports proliferation in a K562 leukemia cell line
title_fullStr Substitution of histidine 30 by asparagine in manganese superoxide dismutase alters biophysical properties and supports proliferation in a K562 leukemia cell line
title_full_unstemmed Substitution of histidine 30 by asparagine in manganese superoxide dismutase alters biophysical properties and supports proliferation in a K562 leukemia cell line
title_short Substitution of histidine 30 by asparagine in manganese superoxide dismutase alters biophysical properties and supports proliferation in a K562 leukemia cell line
title_sort substitution of histidine 30 by asparagine in manganese superoxide dismutase alters biophysical properties and supports proliferation in a k562 leukemia cell line
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8190026/
https://www.ncbi.nlm.nih.gov/pubmed/34021366
http://dx.doi.org/10.1007/s00249-021-01544-2
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