Anion–Induced Amyloid Fibrillation of Human Insulin In vitro

Abstract

AbstractIn the present study, we have focused on the effect of three biologically important salt anions on insulin aggregation. The result of the present study reveals that salts differing in their anions (NaI, NaOAc, and NaNO3) induce the self–assembly formation of insulin even at low physiological (in the micromolar range) salt concentration with efficacy that follows the order I−>CH3COO−>NO3−. Here, the anion–driven aggregation of insulin does not follow either the Hofmeister series or electroselectivity series at very low salt concentrations; instead, the binding of anions at low pH to the positively charged residues of insulin is determined by a mechanism where the salt anions promote the fibrillation of insulin and modify the morphology of the monomeric precursor molecule. Both the nucleation and fibril elongation are controlled by the electrostatic forces and hydrophobic interactions. Aggregation mechanism and aggregate morphology were investigated employing a combination of Thioflavin T (ThT) and ANS fluorescence, circular dichroism (CD), dynamic light scattering (DLS), and transmission electron microscopy (TEM). Overall, the present data will enhance the idea to rationalize the anion effects on the aggregation of amyloid‐prone protein and to understand the influence of charged biomolecules in cellular compartments.