Publication
Article Metrics
Citations
Online attention
Purification and structural characterization of aggregation-prone human TDP-43 involved in neurodegenerative diseases
DOI:
10.1016/j.isci.2020.101159
Authors:
Gareth S. A.
Wright
(University of Liverpool)
,
Tatiana F.
Watanabe
(University of Liverpool)
,
Kangsa
Amporndanai
(University of Liverpool)
,
Steven S.
Plotkin
(University of British Columbia)
,
Neil R.
Cashman
(University of British Columbia)
,
Svetlana V.
Antonyuk
(University of Liverpool)
,
S. Samar
Hasnain
(University of Liverpool)
Co-authored by industrial partner:
No
Type:
Journal Paper
Journal:
Iscience
State:
Published (Approved)
Published:
May 2020

Abstract: Mislocalisation, 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 characterisation has proven 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 NTD domain (residues 1-80) with molecules arranged as parallel spirals with neighbouring molecules arranged in head-to-tail fashion. In order to obtain detergent free, full-length TDP-43 we mutated all six tryptophan residues that provided low-resolution structure of full-length TDP-43 by small angle X-ray scattering and refining the relative positions of individual domains and intrinsically disordered regions in the model using molecular dynamics.
Subject Areas:
Biology and Bio-materials
Instruments:
B21-High Throughput SAXS
,
I03-Macromolecular Crystallography
Added On:
20/05/2020 08:43
Documents:
1-s2.0-S2589004220303448-main.pdf
Discipline Tags:
Neurodegenerative Diseases
Non-Communicable Diseases
Health & Wellbeing
Neurology
Structural biology
Biophysics
Life Sciences & Biotech
Technical Tags:
Diffraction
Scattering
Macromolecular Crystallography (MX)
Small Angle X-ray Scattering (SAXS)