Supramolecular organization of Cytochrome-C into quantum-dot decorated macromolecular network under pH and thermal stress

Abstract

The holo form of Cytochrome-C which is involved in the electron transfer chain of aerobic and anaerobic respiration remains structurally intact by its complex with heme. However, when a prolonged thermal and pH stress was applied, heme was found to abruptly dissociate from the holo protein, resulting in complete collapse of the three-dimensional functional structure. Interestingly, two distinct structures were formed as the consequence of the dissociation event: (i) A macromolecular amyloid-network formed by the collapsed protein fragments, generated by self-oxidation, and (ii) Fe-containing Quantum-Dots (FeQDs) with 2-3 nm diameter formed by heme reorganization. Further adding to intrigue, the FeQDs were re-adsorbed on the surface of the amyloid network leading to FeQD-decorated macromolecular amyloid matrix. The heme-interactant Met80, constituting the amyloidogenic region, initiates the amylogenic cascade, and gradual exposure of Trp59 synergistically emit intrinsic fluorescence alongside FeQDs. The development of the aforementioned events were probed through a multitude of biophysical, chemical and computational analyses like ThT/ANS/intrinsic fluorescence assays, CD-spectroscopy, FETEM/STEM/elemental mapping, Foldamyloid/Foldunfold/Isunstruct/H-protection/LIGplot analyses, etc. The FeQD-decorated amyloid-network was found to exhibit gel-like property, which supported the growth of BHK-21 fibroblast without cytotoxicity. Further studies on FeQD-decorated Cytochrome C amyloid network might open possibilities to design advanced biomaterial for diverse biological applications.

Keywords: Amyloid network; Cytochrome C; FeQDs; Heme; Supramolecular organization.