Issue 36, 2023

A robust yet simple method to generate fluorescent amyloid nanofibers

Abstract

Covalent tagging of fluorophores is central to the mechanistic understanding of important biological processes including protein–protein interaction and protein aggregation. Hence, studies on fluorophore-tagged peptides help in elucidating the molecular mechanism of amyloidogenesis, its cellular internalization, and crosstalk potential. Despite the many advantages the covalently tagged proteins offer, difficulties such as expensive and tedious synthesis and purification protocols have become a matter of concern. Importantly, covalently tagged fluorophores could introduce structural constraints, which may influence the conformation of the monomeric and aggregated forms of proteins. Here, we describe a robust-yet-simple method to make fluorescent-amyloid nanofibers through a coassembly-reaction route that does not alter the aggregation kinetics and the characteristic β-sheet-conformers of resultant nanofibers. Fluorescent amyloid nanofibers derived from insulin, lysozyme, Aβ1–42, and metabolites were successfully fabricated in our study. Importantly, the incorporated fluorophores exhibited remarkable stability, remaining intact without leaching even after undergoing serial dilutions and prolonged storage periods. This method enables monitoring of cellular internalization of the fluorescent-amyloid-nanofibers and the detection of FRET-signals during interfibrillar interactions. This simple and affordable protocol may significantly help amyloid researchers working on both in vitro and animal models.

Graphical abstract: A robust yet simple method to generate fluorescent amyloid nanofibers

  • This article is part of the themed collection: #MyFirstJMCB

Supplementary files

Article information

Article type
Paper
Submitted
26 Cax 2023
Accepted
31 Qad 2023
First published
01 Leq 2023

J. Mater. Chem. B, 2023,11, 8765-8774

A robust yet simple method to generate fluorescent amyloid nanofibers

K. P. Prajapati, M. Ansari, D. K. Yadav, S. Mittal, B. G. Anand and K. Kar, J. Mater. Chem. B, 2023, 11, 8765 DOI: 10.1039/D3TB01203D

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