Versatile Trityl Spin Labels for Nanometer Distance Measurements on Biomolecules In Vitro and within Cells

J. Jacques Jassoy; Andreas Berndhäuser; Fraser Duthie; Sebastian P. Kühn; Gregor Hagelueken; Olav Schiemann

doi:10.1002/anie.201609085

Versatile Trityl Spin Labels for Nanometer Distance Measurements on Biomolecules In Vitro and within Cells

J. Jacques Jassoy, Andreas Berndhäuser, Fraser Duthie, Sebastian P. Kühn, Gregor Hagelueken, Olav Schiemann

University of Bonn

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

109 Scopus citations

Abstract

Structure determination of biomacromolecules under in-cell conditions is a relevant yet challenging task. Electron paramagnetic resonance (EPR) distance measurements in combination with site-directed spin labeling (SDSL) are a valuable tool in this endeavor but the usually used nitroxide spin labels are not well-suited for in-cell measurements. In contrast, triarylmethyl (trityl) radicals are highly persistent, exhibit a long relaxation time and a narrow spectral width. Here, the synthesis of a versatile collection of trityl spin labels and their application in in vitro and in-cell trityl–iron distance measurements on a cytochrome P450 protein are described. The trityl labels show similar labeling efficiencies and better signal-to-noise ratios (SNR) as compared to the popular methanethiosulfonate spin label (MTSSL) and enabled a successful in-cell measurement.

Original language	English
Pages (from-to)	177-181
Number of pages	5
Journal	Angewandte Chemie International Edition in English
Volume	56
Issue number	1
DOIs	https://doi.org/10.1002/anie.201609085
State	Published - 2 Jan 2017
Externally published	Yes

Keywords

EPR spectroscopy
distance measurements
in-cell spectroscopy
protein structures
spin labeling

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10.1002/anie.201609085

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title = "Versatile Trityl Spin Labels for Nanometer Distance Measurements on Biomolecules In Vitro and within Cells",

abstract = "Structure determination of biomacromolecules under in-cell conditions is a relevant yet challenging task. Electron paramagnetic resonance (EPR) distance measurements in combination with site-directed spin labeling (SDSL) are a valuable tool in this endeavor but the usually used nitroxide spin labels are not well-suited for in-cell measurements. In contrast, triarylmethyl (trityl) radicals are highly persistent, exhibit a long relaxation time and a narrow spectral width. Here, the synthesis of a versatile collection of trityl spin labels and their application in in vitro and in-cell trityl–iron distance measurements on a cytochrome P450 protein are described. The trityl labels show similar labeling efficiencies and better signal-to-noise ratios (SNR) as compared to the popular methanethiosulfonate spin label (MTSSL) and enabled a successful in-cell measurement.",

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AU - Jassoy, J. Jacques

AU - Berndhäuser, Andreas

AU - Duthie, Fraser

AU - Kühn, Sebastian P.

AU - Hagelueken, Gregor

AU - Schiemann, Olav

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AB - Structure determination of biomacromolecules under in-cell conditions is a relevant yet challenging task. Electron paramagnetic resonance (EPR) distance measurements in combination with site-directed spin labeling (SDSL) are a valuable tool in this endeavor but the usually used nitroxide spin labels are not well-suited for in-cell measurements. In contrast, triarylmethyl (trityl) radicals are highly persistent, exhibit a long relaxation time and a narrow spectral width. Here, the synthesis of a versatile collection of trityl spin labels and their application in in vitro and in-cell trityl–iron distance measurements on a cytochrome P450 protein are described. The trityl labels show similar labeling efficiencies and better signal-to-noise ratios (SNR) as compared to the popular methanethiosulfonate spin label (MTSSL) and enabled a successful in-cell measurement.

KW - EPR spectroscopy

KW - distance measurements

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ER -

Versatile Trityl Spin Labels for Nanometer Distance Measurements on Biomolecules In Vitro and within Cells

Abstract

Keywords

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