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Semisynthetic Modification of Tau Protein with Di-Ubiquitin Chains for Aggregation Studies

Ubiquitin, a protein modifier that regulates diverse essential cellular processes, is also a component of the protein inclusions characteristic of many neurodegenerative disorders. In Alzheimer’s disease, the microtubule associated tau protein accumulates within damaged neurons in the form of cross-...

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Autores principales: Munari, Francesca, Barracchia, Carlo Giorgio, Parolini, Francesca, Tira, Roberto, Bubacco, Luigi, Assfalg, Michael, D’Onofrio, Mariapina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7352214/
https://www.ncbi.nlm.nih.gov/pubmed/32575755
http://dx.doi.org/10.3390/ijms21124400
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author Munari, Francesca
Barracchia, Carlo Giorgio
Parolini, Francesca
Tira, Roberto
Bubacco, Luigi
Assfalg, Michael
D’Onofrio, Mariapina
author_facet Munari, Francesca
Barracchia, Carlo Giorgio
Parolini, Francesca
Tira, Roberto
Bubacco, Luigi
Assfalg, Michael
D’Onofrio, Mariapina
author_sort Munari, Francesca
collection PubMed
description Ubiquitin, a protein modifier that regulates diverse essential cellular processes, is also a component of the protein inclusions characteristic of many neurodegenerative disorders. In Alzheimer’s disease, the microtubule associated tau protein accumulates within damaged neurons in the form of cross-beta structured filaments. Both mono- and polyubiquitin were found linked to several lysine residues belonging to the region of tau protein that forms the structured core of the filaments. Thus, besides priming the substrate protein for proteasomal degradation, ubiquitin could also contribute to the assembly and stabilization of tau protein filaments. To advance our understanding of the impact of ubiquitination on tau protein aggregation and function, we applied disulfide-coupling chemistry to modify tau protein at position 353 with Lys48- or Lys63-linked di-ubiquitin, two representative polyubiquitin chains that differ in topology and structure. Aggregation kinetics experiments performed on these conjugates reveal that di-ubiquitination retards filament formation and perturbs the fibril elongation rate more than mono-ubiquitination. We further show that di-ubiquitination modulates tau-mediated microtubule assembly. The effects on tau protein aggregation and microtubule polymerization are essentially independent from polyubiquitin chain topology. Altogether, our findings provide novel insight into the consequences of ubiquitination on the functional activity and disease-related behavior of tau protein.
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spelling pubmed-73522142020-07-21 Semisynthetic Modification of Tau Protein with Di-Ubiquitin Chains for Aggregation Studies Munari, Francesca Barracchia, Carlo Giorgio Parolini, Francesca Tira, Roberto Bubacco, Luigi Assfalg, Michael D’Onofrio, Mariapina Int J Mol Sci Article Ubiquitin, a protein modifier that regulates diverse essential cellular processes, is also a component of the protein inclusions characteristic of many neurodegenerative disorders. In Alzheimer’s disease, the microtubule associated tau protein accumulates within damaged neurons in the form of cross-beta structured filaments. Both mono- and polyubiquitin were found linked to several lysine residues belonging to the region of tau protein that forms the structured core of the filaments. Thus, besides priming the substrate protein for proteasomal degradation, ubiquitin could also contribute to the assembly and stabilization of tau protein filaments. To advance our understanding of the impact of ubiquitination on tau protein aggregation and function, we applied disulfide-coupling chemistry to modify tau protein at position 353 with Lys48- or Lys63-linked di-ubiquitin, two representative polyubiquitin chains that differ in topology and structure. Aggregation kinetics experiments performed on these conjugates reveal that di-ubiquitination retards filament formation and perturbs the fibril elongation rate more than mono-ubiquitination. We further show that di-ubiquitination modulates tau-mediated microtubule assembly. The effects on tau protein aggregation and microtubule polymerization are essentially independent from polyubiquitin chain topology. Altogether, our findings provide novel insight into the consequences of ubiquitination on the functional activity and disease-related behavior of tau protein. MDPI 2020-06-20 /pmc/articles/PMC7352214/ /pubmed/32575755 http://dx.doi.org/10.3390/ijms21124400 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Munari, Francesca
Barracchia, Carlo Giorgio
Parolini, Francesca
Tira, Roberto
Bubacco, Luigi
Assfalg, Michael
D’Onofrio, Mariapina
Semisynthetic Modification of Tau Protein with Di-Ubiquitin Chains for Aggregation Studies
title Semisynthetic Modification of Tau Protein with Di-Ubiquitin Chains for Aggregation Studies
title_full Semisynthetic Modification of Tau Protein with Di-Ubiquitin Chains for Aggregation Studies
title_fullStr Semisynthetic Modification of Tau Protein with Di-Ubiquitin Chains for Aggregation Studies
title_full_unstemmed Semisynthetic Modification of Tau Protein with Di-Ubiquitin Chains for Aggregation Studies
title_short Semisynthetic Modification of Tau Protein with Di-Ubiquitin Chains for Aggregation Studies
title_sort semisynthetic modification of tau protein with di-ubiquitin chains for aggregation studies
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7352214/
https://www.ncbi.nlm.nih.gov/pubmed/32575755
http://dx.doi.org/10.3390/ijms21124400
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