Stabilizing Proteins to Prevent Conformational Changes Required for Amyloid Fibril Formation
amyloid, human insulin, protein stability, transmission electron microscopy (TEM), thioflavin T (ThT) assay
Digital Object Identifier (DOI)
Amyloid fibrillation is associated with several human maladies, such as Alzheimer’s, Parkinson’s, Huntington’s diseases, prions, amyotrophic lateral sclerosis, and type 2 diabetes diseases. Gaining insights into the mechanism of amyloid fibril formation and exploring novel approaches to fibrillation inhibition are crucial for preventing amyloid diseases. Here, we hypothesized that ligands capable of stabilizing the native state of query proteins might prevent protein unfolding, which, in turn, may reduce the propensity of proteins to form amyloid fibrils. We demonstrated the efficient inhibition of amyloid formation of the human serum albumin (HSA) (up to 85%) and human insulin (up to 80%) by a nonsteroidal anti-inflammatory drug, ibuprofen (IBFN). IBFN significantly increases the conformational stability of both HSA and insulin, as confirmed by differential scanning calorimetry (DSC). Moreover, increasing concentration of IBFN boosts its amyloid inhibitory propensity in a linear fashion by influencing the nucleation phase as assayed by thioflavin T fluorescence, transmission electron microscopy, and dynamic light scattering. Furthermore, circular dichroism analysis supported the DSC results, showing that IBFN binds to the native state of proteins and almost completely prevents their tendency to lose secondary and tertiary structures. Cell toxicity assay confirms that species formed in the presence of IBFN are less toxic to neuronal cells (SH-SY5Y). These results demonstrate the feasibility of using a small molecule to stabilize the native state of proteins, thereby preventing the amyloidogenic conformational changes, which appear to be the common link in several human amyloid diseases.
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Citation / Publisher Attribution
Journal of Cellular Biochemistry, v. 120, issue 2, p. 2642-2656
Scholar Commons Citation
Siddiqi, Mohammad Khursheed; Alam, Parvez; Malik, Sadia; Majid, Nabeela; Chaturvedi, Sumit Kumar; Rajan, Sudeepa; Ajmal, Mohd Rehan; Khan, Mohsin Vahid; Uversky, Vladimir N.; and Khan, Rizwan Hasan, "Stabilizing Proteins to Prevent Conformational Changes Required for Amyloid Fibril Formation" (2019). Molecular Medicine Faculty Publications. 133.