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In vitro characterization of six STUB1 variants in spinocerebellar ataxia 16 reveals altered structural properties for the encoded CHIP proteins

Spinocerebellar ataxia, autosomal recessive 16 (SCAR16) is caused by biallelic mutations in the STIP1 homology and U-box containing protein 1 (STUB1) gene encoding the ubiquitin E3 ligase and dimeric co-chaperone C-terminus of Hsc70-interacting protein (CHIP). It has been proposed that the disease m...

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Autores principales: Pakdaman, Yasaman, Sanchez-Guixé, Monica, Kleppe, Rune, Erdal, Sigrid, Bustad, Helene J., Bjørkhaug, Lise, Haugarvoll, Kristoffer, Tzoulis, Charalampos, Heimdal, Ketil, Knappskog, Per M., Johansson, Stefan, Aukrust, Ingvild
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5408658/
https://www.ncbi.nlm.nih.gov/pubmed/28396517
http://dx.doi.org/10.1042/BSR20170251
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author Pakdaman, Yasaman
Sanchez-Guixé, Monica
Kleppe, Rune
Erdal, Sigrid
Bustad, Helene J.
Bjørkhaug, Lise
Haugarvoll, Kristoffer
Tzoulis, Charalampos
Heimdal, Ketil
Knappskog, Per M.
Johansson, Stefan
Aukrust, Ingvild
author_facet Pakdaman, Yasaman
Sanchez-Guixé, Monica
Kleppe, Rune
Erdal, Sigrid
Bustad, Helene J.
Bjørkhaug, Lise
Haugarvoll, Kristoffer
Tzoulis, Charalampos
Heimdal, Ketil
Knappskog, Per M.
Johansson, Stefan
Aukrust, Ingvild
author_sort Pakdaman, Yasaman
collection PubMed
description Spinocerebellar ataxia, autosomal recessive 16 (SCAR16) is caused by biallelic mutations in the STIP1 homology and U-box containing protein 1 (STUB1) gene encoding the ubiquitin E3 ligase and dimeric co-chaperone C-terminus of Hsc70-interacting protein (CHIP). It has been proposed that the disease mechanism is related to CHIP’s impaired E3 ubiquitin ligase properties and/or interaction with its chaperones. However, there is limited knowledge on how these mutations affect the stability, folding, and protein structure of CHIP itself. To gain further insight, six previously reported pathogenic STUB1 variants (E28K, N65S, K145Q, M211I, S236T, and T246M) were expressed as recombinant proteins and studied using limited proteolysis, size-exclusion chromatography (SEC), and circular dichroism (CD). Our results reveal that N65S shows increased CHIP dimerization, higher levels of α-helical content, and decreased degradation rate compared with wild-type (WT) CHIP. By contrast, T246M demonstrates a strong tendency for aggregation, a more flexible protein structure, decreased levels of α-helical structures, and increased degradation rate compared with WT CHIP. E28K, K145Q, M211I, and S236T also show defects on structural properties compared with WT CHIP, although less profound than what observed for N65S and T246M. In conclusion, our results illustrate that some STUB1 mutations known to cause recessive SCAR16 have a profound impact on the protein structure, stability, and ability of CHIP to dimerize in vitro. These results add to the growing understanding on the mechanisms behind the disorder.
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spelling pubmed-54086582017-05-04 In vitro characterization of six STUB1 variants in spinocerebellar ataxia 16 reveals altered structural properties for the encoded CHIP proteins Pakdaman, Yasaman Sanchez-Guixé, Monica Kleppe, Rune Erdal, Sigrid Bustad, Helene J. Bjørkhaug, Lise Haugarvoll, Kristoffer Tzoulis, Charalampos Heimdal, Ketil Knappskog, Per M. Johansson, Stefan Aukrust, Ingvild Biosci Rep Research Articles Spinocerebellar ataxia, autosomal recessive 16 (SCAR16) is caused by biallelic mutations in the STIP1 homology and U-box containing protein 1 (STUB1) gene encoding the ubiquitin E3 ligase and dimeric co-chaperone C-terminus of Hsc70-interacting protein (CHIP). It has been proposed that the disease mechanism is related to CHIP’s impaired E3 ubiquitin ligase properties and/or interaction with its chaperones. However, there is limited knowledge on how these mutations affect the stability, folding, and protein structure of CHIP itself. To gain further insight, six previously reported pathogenic STUB1 variants (E28K, N65S, K145Q, M211I, S236T, and T246M) were expressed as recombinant proteins and studied using limited proteolysis, size-exclusion chromatography (SEC), and circular dichroism (CD). Our results reveal that N65S shows increased CHIP dimerization, higher levels of α-helical content, and decreased degradation rate compared with wild-type (WT) CHIP. By contrast, T246M demonstrates a strong tendency for aggregation, a more flexible protein structure, decreased levels of α-helical structures, and increased degradation rate compared with WT CHIP. E28K, K145Q, M211I, and S236T also show defects on structural properties compared with WT CHIP, although less profound than what observed for N65S and T246M. In conclusion, our results illustrate that some STUB1 mutations known to cause recessive SCAR16 have a profound impact on the protein structure, stability, and ability of CHIP to dimerize in vitro. These results add to the growing understanding on the mechanisms behind the disorder. Portland Press Ltd. 2017-04-28 /pmc/articles/PMC5408658/ /pubmed/28396517 http://dx.doi.org/10.1042/BSR20170251 Text en © 2017 The Author(s). http://creativecommons.org/licenses/by/4.0/This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY) (http://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Articles
Pakdaman, Yasaman
Sanchez-Guixé, Monica
Kleppe, Rune
Erdal, Sigrid
Bustad, Helene J.
Bjørkhaug, Lise
Haugarvoll, Kristoffer
Tzoulis, Charalampos
Heimdal, Ketil
Knappskog, Per M.
Johansson, Stefan
Aukrust, Ingvild
In vitro characterization of six STUB1 variants in spinocerebellar ataxia 16 reveals altered structural properties for the encoded CHIP proteins
title In vitro characterization of six STUB1 variants in spinocerebellar ataxia 16 reveals altered structural properties for the encoded CHIP proteins
title_full In vitro characterization of six STUB1 variants in spinocerebellar ataxia 16 reveals altered structural properties for the encoded CHIP proteins
title_fullStr In vitro characterization of six STUB1 variants in spinocerebellar ataxia 16 reveals altered structural properties for the encoded CHIP proteins
title_full_unstemmed In vitro characterization of six STUB1 variants in spinocerebellar ataxia 16 reveals altered structural properties for the encoded CHIP proteins
title_short In vitro characterization of six STUB1 variants in spinocerebellar ataxia 16 reveals altered structural properties for the encoded CHIP proteins
title_sort in vitro characterization of six stub1 variants in spinocerebellar ataxia 16 reveals altered structural properties for the encoded chip proteins
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5408658/
https://www.ncbi.nlm.nih.gov/pubmed/28396517
http://dx.doi.org/10.1042/BSR20170251
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