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| Main Authors: | , , , , , |
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| Format: | Artículo Open Access |
| Published: |
Wiley
2024
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| Subjects: | |
| Online Access: | https://onlinelibrary.wiley.com/doi/10.1111/php.14004 |
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Table of Contents:
- Tracking heterogenous protein aggregation at nanoscale through fluorescence correlation spectroscopy Bisal Halder Shreya Ghosh Tanmoy Khan Subhendu Pal Nilimesh Das Pratik Sen Photochemistry and Photobiology AbstractVarious biophysical techniques have been extensively employed to study protein aggregation due to its significance. Traditionally, these methods detect aggregation at micrometer length scales and micromolar concentrations. However, unlike in vitro, protein aggregation typically occurs at nanomolar concentrations in vivo. Here, using fluorescence correlation spectroscopy (FCS), we captured bromelain aggregation at concentrations as low as ~20 nM, surpassing the detection limit of traditional methods like thioflavin T fluorescence, scattering, and fluorescence microscopy by more than one order of magnitude. Moreover, using thioflavin T fluorescence‐based FCS, we have detected larger aggregates at higher bromelain concentrations, which is undetectable in FCS otherwise. Importantly, our study reveals inherent heterogeneity in bromelain aggregation, inaccessible to ensemble‐averaged techniques. The presented report may provide a platform for the characterization of premature aggregates at very low protein concentrations, which are thought to be functionally significant species in protein aggregation‐induced diseases. 10.1111/php.14004 http://onlinelibrary.wiley.com/termsAndConditions#vor