Stabilizing proteins to prevent conformational changes required for amyloid fibril formation

Area of Science:

• Biochemistry
• Molecular Biology
• Neuroscience

Background:

• Amyloid fibrillation is implicated in numerous human diseases, including Alzheimer's, Parkinson's, and type 2 diabetes.
• Understanding amyloid fibril formation mechanisms and developing inhibitors are critical for disease prevention.

Purpose of the Study:

• To investigate if stabilizing the native state of proteins can inhibit amyloid fibril formation.
• To evaluate ibuprofen (IBFN) as a potential inhibitor of amyloidogenesis.

Main Methods:

• Differential scanning calorimetry (DSC) and circular dichroism (CD) to assess protein conformational stability.
• Thioflavin T fluorescence, transmission electron microscopy (TEM), and dynamic light scattering (DLS) to monitor amyloid formation.
• Cell toxicity assays using neuronal cells (SH-SY5Y).

Main Results:

• Ibuprofen (IBFN) inhibited amyloid formation of human serum albumin (HSA) by up to 85% and human insulin by up to 80%.
• IBFN increased the conformational stability of HSA and insulin, binding to their native states and preventing structural changes.
• IBFN-treated amyloid species exhibited reduced toxicity to neuronal cells.

Conclusions:

• Stabilizing the native state of proteins with small molecules like ibuprofen is a feasible strategy to prevent amyloidogenic conformational changes.
• This approach holds promise for developing novel therapeutic strategies against a range of amyloid-related diseases.