Measuring Nanometer Distances in Proteins and Rigid Rulers between 19F and Gd3+ by Integration of 19F-ENDOR Signal Intensities

Martyna Judd; Mian Qi; Elwy H. Abdelkader; Haocheng Qianzhu; Anton Savitsky; Thomas Huber; Jeffrey R. Harmer; Adelheid Godt; Gottfried Otting; Nicholas Cox

doi:10.1021/jacs.4c13530

Measuring Nanometer Distances in Proteins and Rigid Rulers between ¹⁹F and Gd³⁺ by Integration of ¹⁹F-ENDOR Signal Intensities

Martyna Judd^*, Mian Qi, Elwy H. Abdelkader, Haocheng Qianzhu, Anton Savitsky, Thomas Huber, Jeffrey R. Harmer, Adelheid Godt, Gottfried Otting, Nicholas Cox^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

4 Citations (Scopus)

Abstract

¹⁹F ENDOR is emerging as a powerful tool in structural biology for measuring distances in proteins labeled with ¹⁹F and a paramagnetic tag. Due to spin-spin relaxation and line width limitations, it has been difficult to determine intertag distances larger than about 15 Å. Using a set of geometrically well-defined rulers and spin-labeled proteins, we show that ¹⁹F-Gd³⁺ distances up to 20 Å can be accessed by integrating the intensity of the ENDOR spectrum, with distances approaching 30 Å potentially in reach as well. This method is robust when the intensities are scaled to a known reference, and provides scope for nanometer-scale triangulation of the coordinates of a ligand in a protein-ligand complex.

Original language	English
Pages (from-to)	16826–16835
Number of pages	10
Journal	Journal of the American Chemical Society
Volume	147
Issue number	20
DOIs	https://doi.org/10.1021/jacs.4c13530
Publication status	Published - 21 May 2025

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title = "Measuring Nanometer Distances in Proteins and Rigid Rulers between 19F and Gd3+ by Integration of 19F-ENDOR Signal Intensities",

abstract = "19F ENDOR is emerging as a powerful tool in structural biology for measuring distances in proteins labeled with 19F and a paramagnetic tag. Due to spin-spin relaxation and line width limitations, it has been difficult to determine intertag distances larger than about 15 {\AA}. Using a set of geometrically well-defined rulers and spin-labeled proteins, we show that 19F-Gd3+ distances up to 20 {\AA} can be accessed by integrating the intensity of the ENDOR spectrum, with distances approaching 30 {\AA} potentially in reach as well. This method is robust when the intensities are scaled to a known reference, and provides scope for nanometer-scale triangulation of the coordinates of a ligand in a protein-ligand complex.",

author = "Martyna Judd and Mian Qi and Abdelkader, \{Elwy H.\} and Haocheng Qianzhu and Anton Savitsky and Thomas Huber and Harmer, \{Jeffrey R.\} and Adelheid Godt and Gottfried Otting and Nicholas Cox",

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day = "21",

doi = "10.1021/jacs.4c13530",

language = "English",

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T1 - Measuring Nanometer Distances in Proteins and Rigid Rulers between 19F and Gd3+ by Integration of 19F-ENDOR Signal Intensities

AU - Judd, Martyna

AU - Qi, Mian

AU - Abdelkader, Elwy H.

AU - Qianzhu, Haocheng

AU - Savitsky, Anton

AU - Huber, Thomas

AU - Harmer, Jeffrey R.

AU - Godt, Adelheid

AU - Otting, Gottfried

AU - Cox, Nicholas

PY - 2025/5/21

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N2 - 19F ENDOR is emerging as a powerful tool in structural biology for measuring distances in proteins labeled with 19F and a paramagnetic tag. Due to spin-spin relaxation and line width limitations, it has been difficult to determine intertag distances larger than about 15 Å. Using a set of geometrically well-defined rulers and spin-labeled proteins, we show that 19F-Gd3+ distances up to 20 Å can be accessed by integrating the intensity of the ENDOR spectrum, with distances approaching 30 Å potentially in reach as well. This method is robust when the intensities are scaled to a known reference, and provides scope for nanometer-scale triangulation of the coordinates of a ligand in a protein-ligand complex.

AB - 19F ENDOR is emerging as a powerful tool in structural biology for measuring distances in proteins labeled with 19F and a paramagnetic tag. Due to spin-spin relaxation and line width limitations, it has been difficult to determine intertag distances larger than about 15 Å. Using a set of geometrically well-defined rulers and spin-labeled proteins, we show that 19F-Gd3+ distances up to 20 Å can be accessed by integrating the intensity of the ENDOR spectrum, with distances approaching 30 Å potentially in reach as well. This method is robust when the intensities are scaled to a known reference, and provides scope for nanometer-scale triangulation of the coordinates of a ligand in a protein-ligand complex.

UR - https://www.scopus.com/pages/publications/105004348835

U2 - 10.1021/jacs.4c13530

DO - 10.1021/jacs.4c13530

M3 - Article

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JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

IS - 20

ER -

Measuring Nanometer Distances in Proteins and Rigid Rulers between ¹⁹F and Gd³⁺ by Integration of ¹⁹F-ENDOR Signal Intensities

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