@article{4b5480990ea545399d2b241a5cf01095,
title = "Triggered Functional Dynamics of AsLOV2 by Time-Resolved Electron Paramagnetic Resonance at High Magnetic Fields",
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.",
keywords = "EPR Spectroscopy, Gadolinium, Protein Structures, Tigger, Time-Resolved Spectroscopy",
author = "Shiny Maity and Price, {Brad D.} and Wilson, {C. Blake} and Arnab Mukherjee and Matthieu Starck and David Parker and Wilson, {Maxwell Z.} and Lovett, {Janet E.} and Songi Han and Sherwin, {Mark S.}",
note = "Funding Information: The authors would like to thank Karen Tsay for her thoughtful insight and assistance in running Q‐band DEER experiments. We would also like to acknowledge support from the NSF through grant MCB 2025860 and the UC Office of the President Multicampus Research Programs and Initiatives under MRI‐19‐601107 for the development of TiGGER and the core research presented here. Biochemical and complementary biophysical studies of AsLOV2 were supported by the NIH MIRA through grant R35GM136411. The authors also acknowledge primary funding support from the US Army Research Office under Cooperative Agreement Numbers W911NF‐19‐2‐0026 and Task 23 ICB‐2019‐BEM‐01 for the Institute for Collaborative Biotechnologies. J.E.L. thanks The Royal Society for a University Research Fellowship. Figures were created in Python using SciencePlots. All data and programs used for this publication are available at Ref. [78] and Ref. [79] respectively. [77] Publisher Copyright: {\textcopyright} 2023 Wiley-VCH GmbH.",
year = "2023",
month = mar,
day = "20",
doi = "10.1002/anie.202212832",
language = "English",
volume = "62",
journal = "Angewandte Chemie. International Edition",
issn = "1433-7851",
publisher = "John Wiley & Sons Ltd.",
number = "13",
}