Wright, GSA ORCID: 0000-0002-3756-9634, Watanabe, TF, Amporndanai, K, Plotkin, SS, Cashman, NR, Antonyuk, SV ORCID: 0000-0002-2779-9946 and Hasnain, SS
(2020)
Purification and Structural Characterization of Aggregation-Prone Human TDP-43 Involved in Neurodegenerative Diseases.
iScience, 23 (6).
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Abstract
© 2020 The Author(s) Mislocalization, cleavage, and aggregation of the human protein TDP-43 is found in many neurodegenerative diseases. As is the case with many other proteins that are completely or partially structurally disordered, production of full-length recombinant TDP-43 in the quantities necessary for structural characterization has proved difficult. We show that the full-length TDP-43 protein and two truncated N-terminal constructs 1-270 and 1-263 can be heterologously expressed in E. coli. Full-length TDP-43 could be prevented from aggregation during purification using a detergent. Crystals grown from an N-terminal construct (1-270) revealed only the N-terminal domain (residues 1-80) with molecules arranged as parallel spirals with neighboring molecules arranged in head-to-tail fashion. To obtain detergent-free, full-length TDP-43 we mutated all six tryptophan residues to alanine. This provided sufficient soluble protein to collect small-angle X-ray scattering data. Refining relative positions of individual domains and intrinsically disordered regions against this data yielded a model of full-length TDP-43.
Item Type: | Article |
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Depositing User: | Symplectic Admin |
Date Deposited: | 03 Jun 2020 09:43 |
Last Modified: | 18 Jan 2023 23:50 |
DOI: | 10.1016/j.isci.2020.101159 |
Open Access URL: | https://www.sciencedirect.com/science/article/pii/... |
URI: | https://livrepository.liverpool.ac.uk/id/eprint/3089400 |
Available Versions of this Item
- Purification and Structural Characterization of Aggregation-Prone Human TDP-43 Involved in Neurodegenerative Diseases. (deposited 03 Jun 2020 09:43) [Currently Displayed]