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Genetic, structural, and functional analysis of pathogenic variations causing methylmalonyl-CoA epimerase deficiency

Human methylmalonyl-CoA epimerase (MCEE) catalyzes the interconversion of d-methylmalonyl-CoA and l-methylmalonyl-CoA in propionate catabolism. Autosomal recessive pathogenic variations in MCEE reportedly cause methylmalonic aciduria (MMAuria) in eleven patients. We investigated a cohort of 150 indi...

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Autores principales: Heuberger, Kathrin, Bailey, Henry J., Burda, Patricie, Chaikuad, Apirat, Krysztofinska, Ewelina, Suormala, Terttu, Bürer, Céline, Lutz, Seraina, Fowler, Brian, Froese, D. Sean, Yue, Wyatt W., Baumgartner, Matthias R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6525113/
https://www.ncbi.nlm.nih.gov/pubmed/30682498
http://dx.doi.org/10.1016/j.bbadis.2019.01.021
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author Heuberger, Kathrin
Bailey, Henry J.
Burda, Patricie
Chaikuad, Apirat
Krysztofinska, Ewelina
Suormala, Terttu
Bürer, Céline
Lutz, Seraina
Fowler, Brian
Froese, D. Sean
Yue, Wyatt W.
Baumgartner, Matthias R.
author_facet Heuberger, Kathrin
Bailey, Henry J.
Burda, Patricie
Chaikuad, Apirat
Krysztofinska, Ewelina
Suormala, Terttu
Bürer, Céline
Lutz, Seraina
Fowler, Brian
Froese, D. Sean
Yue, Wyatt W.
Baumgartner, Matthias R.
author_sort Heuberger, Kathrin
collection PubMed
description Human methylmalonyl-CoA epimerase (MCEE) catalyzes the interconversion of d-methylmalonyl-CoA and l-methylmalonyl-CoA in propionate catabolism. Autosomal recessive pathogenic variations in MCEE reportedly cause methylmalonic aciduria (MMAuria) in eleven patients. We investigated a cohort of 150 individuals suffering from MMAuria of unknown origin, identifying ten new patients with pathogenic variations in MCEE. Nine patients were homozygous for the known nonsense variation p.Arg47* (c.139C > T), and one for the novel missense variation p.Ile53Arg (c.158T > G). To understand better the molecular basis of MCEE deficiency, we mapped p.Ile53Arg, and two previously described pathogenic variations p.Lys60Gln and p.Arg143Cys, onto our 1.8 Å structure of wild-type (wt) human MCEE. This revealed potential dimeric assembly disruption by p.Ile53Arg, but no clear defects from p.Lys60Gln or p.Arg143Cys. We solved the structure of MCEE-Arg143Cys to 1.9 Å and found significant disruption of two important loop structures, potentially impacting surface features as well as the active-site pocket. Functional analysis of MCEE-Ile53Arg expressed in a bacterial recombinant system as well as patient-derived fibroblasts revealed nearly undetectable soluble protein levels, defective globular protein behavior, and using a newly developed assay, lack of enzymatic activity - consistent with misfolded protein. By contrast, soluble protein levels, unfolding characteristics and activity of MCEE-Lys60Gln were comparable to wt, leaving unclear how this variation may cause disease. MCEE-Arg143Cys was detectable at comparable levels to wt MCEE, but had slightly altered unfolding kinetics and greatly reduced activity. These studies reveal ten new patients with MCEE deficiency and rationalize misfolding and loss of activity as molecular defects in MCEE-type MMAuria.
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spelling pubmed-65251132019-06-01 Genetic, structural, and functional analysis of pathogenic variations causing methylmalonyl-CoA epimerase deficiency Heuberger, Kathrin Bailey, Henry J. Burda, Patricie Chaikuad, Apirat Krysztofinska, Ewelina Suormala, Terttu Bürer, Céline Lutz, Seraina Fowler, Brian Froese, D. Sean Yue, Wyatt W. Baumgartner, Matthias R. Biochim Biophys Acta Mol Basis Dis Article Human methylmalonyl-CoA epimerase (MCEE) catalyzes the interconversion of d-methylmalonyl-CoA and l-methylmalonyl-CoA in propionate catabolism. Autosomal recessive pathogenic variations in MCEE reportedly cause methylmalonic aciduria (MMAuria) in eleven patients. We investigated a cohort of 150 individuals suffering from MMAuria of unknown origin, identifying ten new patients with pathogenic variations in MCEE. Nine patients were homozygous for the known nonsense variation p.Arg47* (c.139C > T), and one for the novel missense variation p.Ile53Arg (c.158T > G). To understand better the molecular basis of MCEE deficiency, we mapped p.Ile53Arg, and two previously described pathogenic variations p.Lys60Gln and p.Arg143Cys, onto our 1.8 Å structure of wild-type (wt) human MCEE. This revealed potential dimeric assembly disruption by p.Ile53Arg, but no clear defects from p.Lys60Gln or p.Arg143Cys. We solved the structure of MCEE-Arg143Cys to 1.9 Å and found significant disruption of two important loop structures, potentially impacting surface features as well as the active-site pocket. Functional analysis of MCEE-Ile53Arg expressed in a bacterial recombinant system as well as patient-derived fibroblasts revealed nearly undetectable soluble protein levels, defective globular protein behavior, and using a newly developed assay, lack of enzymatic activity - consistent with misfolded protein. By contrast, soluble protein levels, unfolding characteristics and activity of MCEE-Lys60Gln were comparable to wt, leaving unclear how this variation may cause disease. MCEE-Arg143Cys was detectable at comparable levels to wt MCEE, but had slightly altered unfolding kinetics and greatly reduced activity. These studies reveal ten new patients with MCEE deficiency and rationalize misfolding and loss of activity as molecular defects in MCEE-type MMAuria. Elsevier 2019-06-01 /pmc/articles/PMC6525113/ /pubmed/30682498 http://dx.doi.org/10.1016/j.bbadis.2019.01.021 Text en © 2019 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Heuberger, Kathrin
Bailey, Henry J.
Burda, Patricie
Chaikuad, Apirat
Krysztofinska, Ewelina
Suormala, Terttu
Bürer, Céline
Lutz, Seraina
Fowler, Brian
Froese, D. Sean
Yue, Wyatt W.
Baumgartner, Matthias R.
Genetic, structural, and functional analysis of pathogenic variations causing methylmalonyl-CoA epimerase deficiency
title Genetic, structural, and functional analysis of pathogenic variations causing methylmalonyl-CoA epimerase deficiency
title_full Genetic, structural, and functional analysis of pathogenic variations causing methylmalonyl-CoA epimerase deficiency
title_fullStr Genetic, structural, and functional analysis of pathogenic variations causing methylmalonyl-CoA epimerase deficiency
title_full_unstemmed Genetic, structural, and functional analysis of pathogenic variations causing methylmalonyl-CoA epimerase deficiency
title_short Genetic, structural, and functional analysis of pathogenic variations causing methylmalonyl-CoA epimerase deficiency
title_sort genetic, structural, and functional analysis of pathogenic variations causing methylmalonyl-coa epimerase deficiency
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6525113/
https://www.ncbi.nlm.nih.gov/pubmed/30682498
http://dx.doi.org/10.1016/j.bbadis.2019.01.021
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