Research

Medical Sciences

Title :

Investigation of liquid-liquid phase separation associated with heat shock protein70 and their role in regulating neurodegenerative disease

Area of research :

Medical Sciences

Focus area :

Molecular Biology, Neurodegeneration

Principal Investigator :

Dr. Soumit Sankar Mandal, Indian Institute of Science Education and Research (IISER), Tirupati, Andhra Pradesh

Timeline Start Year :

2024

Timeline End Year :

2027

Contact info :

Details

Executive Summary :

Chaperone proteins, such as hsp40, hsp60, hsp70, and small hsps, stabilize unstructured proteins in cells, preventing aggregation. However, under pathological conditions, some proteins, such as intrinsically disordered proteins (IDPs), form unstable protein structures, leading to the formation of toxic amyloid fibrils. These proteins undergo phase separation, forming viscous droplets that mature into insoluble aggregates. These droplets can recruit drugs, peptides, or proteins, which can either accelerate or inhibit aggregation. Inhibiting aggregation is crucial for treatment. This proposal aims to examine the phase separation phenomenon associated with chaperone proteins, which stabilize unstructured polypeptides (UPs) and prevent aggregation. Using hsps with a long unstructured domain, such as hsp70, the disordered region consists of approximately 30-40 amino acids. The study will examine phase separation in the presence of macromolecular crowding agents and optimize protein and salt concentrations. The study will also monitor the recruitment of amyloid peptides into condensates and their aggregation kinetics using Thioflavin T assay, turbidity assay based on spectroscopic techniques, and imaging using confocal microscopy and TEM. The conformational dynamics of chaperone proteins and unstructured polypeptides will be obtained at the molecular level using optical tweezer-based surface tension and viscosity measurement.

Total Budget (INR):

52,65,832

Organizations involved