Purification and Structural Characterization of Aggregation-Prone Human TDP-43 Involved in Neurodegenerative Diseases

Purification and Structural Characterization of Aggregation-Prone Human TDP-43 Involved in Neurodegenerative Diseases

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Summary: 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 tourettebrewing.com 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 usc trojans snapback hat 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.