Monitoring protein conformational changes using fluorescent nanoantennas

@article{Harroun2021MonitoringPC,
  title={Monitoring protein conformational changes using fluorescent nanoantennas},
  author={Scott G. Harroun and Dominic Lauzon and Maximilian Ebert and Arnaud Desrosiers and Xiaomeng Wang and Alexis Vall{\'e}e-B{\'e}lisle},
  journal={Nature Methods},
  year={2021},
  volume={19},
  pages={71-80}
}
Understanding the relationship between protein structural dynamics and function is crucial for both basic research and biotechnology. However, methods for studying the fast dynamics of structural changes are limited. Here, we introduce fluorescent nanoantennas as a spectroscopic technique to sense and report protein conformational changes through noncovalent dye-protein interactions. Using experiments and molecular simulations, we detect and characterize five distinct conformational states of… 
2 Citations

Structural characterization of functionally important chloride binding sites in the marine Vibrio alkaline phosphatase

Results reveal the complex nature and dynamics of chloride binding to enzymes through long-range modulation of electronic potential in the vicinity of the active site, resulting in increased catalytic efficiency and stability.

References

SHOWING 1-10 OF 88 REFERENCES

Distance mapping in proteins using fluorescence spectroscopy: the tryptophan-induced quenching (TrIQ) method.

TrIQ provides a straightforward, readily accessible approach for mapping distances within proteins and monitoring conformational changes using fluorescence spectroscopy, and is complementary to traditional fluorescence resonance energy transfer methods that occur over distances too large for precise studies of protein structure.

Visualizing conformational dynamics of proteins in solution and at the cell membrane

This work combines amber codon suppression to introduce a donor fluorescent noncanonical amino acid with a new, biocompatible approach for labeling proteins with acceptor transition metals in a method called ACCuRET (Anap Cyclen-Cu2+ resonance energy transfer), which measures absolute distances and distance changes with high precision and accuracy.

Harnessing photoinduced electron transfer to optically determine protein sub-nanoscale atomic distances

This work presents an optical approach, distance-encoding photoinduced electron transfer (DEPET), capable of the simultaneous study of protein structure and function, and reports the first implementation of DEPET on human large-conductance K+ channels under voltage clamp.

Visualizing transient protein-folding intermediates by tryptophan-scanning mutagenesis

This work shows that transient states during protein folding can be characterized by measuring the fluorescence of tryptophan residues, introduced at many solvent-exposed positions to determine whether each position is native-like, denatured-like or non-native-like in the intermediate state.

Detection of dihydrofolate reductase conformational change by FRET using two fluorescent amino acids.

It is demonstrated that amino acids containing small fluorophores can be introduced into DHFR with minimal disruption of function and in a fashion that enables sensitive monitoring of changes in DHFR conformation.

Toward dynamic structural biology: Two decades of single-molecule Förster resonance energy transfer

The evolution of smFRET as a key tool for “dynamic structural biology” over the past 22 years is reviewed and the prospects for its use in applications such as biosensing, high-throughput screening, and molecular diagnostics are highlighted.

Initial state of DNA-Dye complex sets the stage for protein induced fluorescence modulation

A perspective on understanding the general phenomenon of induced fluorescence modulation is provided, allowing for proper integration of both PIFE and PIFQ with existing bulk and single-molecule fluorescence techniques.

Investigating tryptophan quenching of fluorescein fluorescence under protolytic equilibrium.

The fluorescence of fluorescein is explored in buffered solutions, in different acidic and basic conditions, and at varied concentrations of tryptophan derivatives, using steady-state absorption and fluorescence spectroscopy, combined with fluorescence lifetime measurements.

Fluorogenic protein labelling: a review of photophysical quench mechanisms and principles of fluorogen design

The mechanistic principles of three major photophysical quenching mechanisms involving Forster resonance energy transfer, through-bond energy transfer (TBET), and photoinduced electron transfer (PeT) are reviewed and design principles for novel fluorogenic probes based on an understanding of these quench mechanisms are presented.
...