Effect of Zinc on the in vitro aggregation and phase separation of TDP-43 protein implicated in Amyotrophic Lateral Sclerosis

S, Preethi and Patel, Basant Kumar (2019) Effect of Zinc on the in vitro aggregation and phase separation of TDP-43 protein implicated in Amyotrophic Lateral Sclerosis. Masters thesis, Indian institute of technology Hyderabad.

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Abstract

Amyotrophic Lateral Sclerosis (ALS) also known as Lou Gehrig’s disease is caused by the degeneration of upper motor neurons leading to paralysis. About 25 potential biomarkers are found to be implicated in ALS. Transactive response DNA binding protein (TDP-43), a RNA/DNA binding protein was found as Tau-negative, ubiquitin-positive cytoplasmic inclusions in ALS patients. Zinc is the second most abundant trace metal found in our body for regulating various biological functions. There are some reports stating that Zinc can also bind to TDP-43 protein and can cause aggregation in vivo. In this study, we have found the possible zinc binding sites that can be present in the TDP-43 protein by using Metal ion binding (MIB) prediction and docking server. As there are some possible zinc binding sites present, they are further examined for their in vitro aggregation with full-length TDP-43, Cterminal fragment TDP-432c (wild type and mutant A315T) by Thioflavin-T and turbidity assays. Further, it was checked for the phase separation by using fluorescence microscopy. These data showed that zinc could accelerate the in vitro aggregation of TDP-43 protein and phase separate it to insoluble, irreversible aggregates.

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