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Triggered Functional Dynamics of AsLOV2 by Time‐Resolved Electron Paramagnetic Resonance at High Magnetic Fields

Maity, Shiny; Price, Brad D.; Wilson, C. Blake; Mukherjee, Arnab; Starck, Matthieu; Parker, David; Wilson, Maxwell Z.; Lovett, Janet E.; Han, Songi; Sherwin, Mark S.

Authors

Shiny Maity

Brad D. Price

C. Blake Wilson

Arnab Mukherjee

Matthieu Starck

Maxwell Z. Wilson

Janet E. Lovett

Songi Han

Mark S. Sherwin



Abstract

We present time-resolved Gd−Gd electron paramagnetic resonance (TiGGER) at 240 GHz for tracking inter-residue distances during a protein's mechanical cycle in the solution state. TiGGER makes use of Gd-sTPATCN spin labels, whose favorable qualities include a spin-7/2 EPR-active center, short linker, narrow intrinsic linewidth, and virtually no anisotropy at high fields (8.6 T) when compared to nitroxide spin labels. Using TiGGER, we determined that upon light activation, the C-terminus and N-terminus of AsLOV2 separate in less than 1 s and relax back to equilibrium with a time constant of approximately 60 s. TiGGER revealed that the light-activated long-range mechanical motion is slowed in the Q513A variant of AsLOV2 and is correlated to the similarly slowed relaxation of the optically excited chromophore as described in recent literature. TiGGER has the potential to valuably complement existing methods for the study of triggered functional dynamics in proteins.

Citation

Maity, S., Price, B. D., Wilson, C. B., Mukherjee, A., Starck, M., Parker, D., …Sherwin, M. S. (2023). Triggered Functional Dynamics of AsLOV2 by Time‐Resolved Electron Paramagnetic Resonance at High Magnetic Fields. Angewandte Chemie International Edition, 62(13), Article e202212832. https://doi.org/10.1002/anie.202212832

Journal Article Type Article
Online Publication Date Mar 14, 2023
Publication Date Mar 20, 2023
Deposit Date May 5, 2023
Publicly Available Date Mar 29, 2024
Journal Angewandte Chemie International Edition
Print ISSN 1433-7851
Electronic ISSN 1521-3773
Publisher Wiley
Peer Reviewed Peer Reviewed
Volume 62
Issue 13
Article Number e202212832
DOI https://doi.org/10.1002/anie.202212832
Public URL https://durham-repository.worktribe.com/output/1175202

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