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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-...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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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. |
format | Online Article Text |
id | pubmed-7352214 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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