Bifunctionality of Iminodiacetic Acid-Modified Lysozyme on Inhibiting Zn²⁺-Mediated Amyloid β-Protein Aggregation

Aggregation of amyloid β-proteins (Aβ) mediated by metal ions such as Zn²⁺ has been suggested to be implicated in the progression of Alzheimer's disease (AD). Hence, development of bifunctional agents capable of inhibiting Aβ aggregation and modulating metal-Aβ species is an effective strategy for the treatment of AD. In this work, we modified iminodiacetic acid (IDA) onto human lysozyme (hLys) surface to create an inhibitor of Zn²⁺-mediated Aβ aggregation and cytotoxicity. The IDA-modified hLys (IDA-hLys) retained the stability and biocompatibility of native hLys. Extensive biophysical and biological analyses indicated that IDA-hLys significantly attenuated Zn²⁺-mediated Aβ aggregation and cytotoxicity due to its strong binding affinity for Zn²⁺, whereas native hLys showed little effect. Stopped-flow fluorescence spectroscopy showed that IDA-hLys could protect Aβ from Zn²⁺-induced aggregation and rapidly depolymerize Zn²⁺-Aβ aggregates. The research indicates that IDA-hLys is a bifunctional agent capable of inhibiting Aβ fibrillization and modulating Zn²⁺-mediated Aβ aggregation and cytotoxicity as a strong Zn²⁺ chelator.