Pulse dipolar EPR for determining nanomolar binding affinities

Katrin Ackermann; Joshua Wort; Bela E. Bode

doi:10.1039/D2CC02360A

Pulse dipolar EPR for determining nanomolar binding affinities

Katrin Ackermann, Joshua Wort, Bela E. Bode^*

^*Corresponding author for this work

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

Abstract

Protein interaction studies often require very low concentrations and highly sensitive biophysical methods. Here, we demonstrate that pulse dipolar electron paramagnetic resonance spectroscopy allows measuring dissociation constants in the nanomolar range. This approach is appealing for concentration-limited biomolecular systems and medium-to-high-affinity binding studies, demonstrated here at 50 nanomolar protein concentration.

Original language	English
Pages (from-to)	8790-8793
Number of pages	4
Journal	Chemical Communications
Volume	58
Issue number	63
Early online date	11 Jul 2022
DOIs	https://doi.org/10.1039/D2CC02360A
Publication status	Published - 14 Aug 2022

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10.1039/D2CC02360ALicence: CC BY

Ackermann_2022_CC_Pulsedipolar_CC
Copyright © 2022 The Author(s). This article is licensed under a Creative Commons Attribution 3.0 Unported Licence.
Final published version, 833 KBLicence: CC BY

5 Finished

ISSF3 - Bela Bode: ISSF3 - Bela Bode
Bode, B. E. (PI)
The Wellcome Trust
1/04/21 → 30/09/21
Project: Standard
Enabling Shaped Pulse Capability: Enabling Shaped Pulse Capability for Superior Biological Structural Determination Using EPR Spectroscopy.
Lovett, J. E. (PI), Bode, B. E. (CoI), Penedo, C. (CoI), Pitt, S. J. (CoI), Schwarz-Linek, U. (CoI), Smith, G. M. (CoI), Watson, A. J. B. (CoI) & White, M. (CoI)
BBSRC
1/10/20 → 30/09/21
Project: Standard
Understanding sensitivity gains in pulse: Understanding sensitivity gains in pulse EPR on multimeric membrane proteins
Bode, B. E. (PI) & Pliotas, C. (CoI)
The Leverhulme Trust
14/05/19 → 18/02/22
Project: Standard

Pulse dipolar EPR for determining nanomolar binding affinities (dataset)
Ackermann, K. (Creator), Wort, J. (Creator) & Bode, B. E. (Creator), University of St Andrews, 13 Jul 2022
DOI: 10.17630/1fece9a1-67d8-42b1-8a46-08944a6ab023
Dataset

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1 Comment/debate

Correction to "Pulse dipolar EPR for determining nanomolar binding affinities": (Chemical Communications (2022), 58:63, DOI:10.1039/d2cc02360a)
Ackermann, K., Wort, J. L. & Bode, B. E., 9 Aug 2022, In: Chemical Communications. 58, 67, p. 9438 1 p.
Research output: Contribution to journal › Comment/debate › peer-review

Open Access

Cite this

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title = "Pulse dipolar EPR for determining nanomolar binding affinities",

abstract = "Protein interaction studies often require very low concentrations and highly sensitive biophysical methods. Here, we demonstrate that pulse dipolar electron paramagnetic resonance spectroscopy allows measuring dissociation constants in the nanomolar range. This approach is appealing for concentration-limited biomolecular systems and medium-to-high-affinity binding studies, demonstrated here at 50 nanomolar protein concentration.",

author = "Katrin Ackermann and Joshua Wort and Bode, {Bela E.}",

note = "This research was funded, in whole or in part, by the Wellcome Trust (204821/Z/16/Z). BEB and KA acknowledge support by the Leverhulme Trust (RPG-2018–397). JLW acknowledges support by the BBSRC DTP Eastbio. BEB acknowledges equipment funding by BBSRC (BB/R013780/1 and BB/T017740/1).",

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T1 - Pulse dipolar EPR for determining nanomolar binding affinities

AU - Ackermann, Katrin

AU - Wort, Joshua

AU - Bode, Bela E.

N1 - This research was funded, in whole or in part, by the Wellcome Trust (204821/Z/16/Z). BEB and KA acknowledge support by the Leverhulme Trust (RPG-2018–397). JLW acknowledges support by the BBSRC DTP Eastbio. BEB acknowledges equipment funding by BBSRC (BB/R013780/1 and BB/T017740/1).

PY - 2022/8/14

Y1 - 2022/8/14

N2 - Protein interaction studies often require very low concentrations and highly sensitive biophysical methods. Here, we demonstrate that pulse dipolar electron paramagnetic resonance spectroscopy allows measuring dissociation constants in the nanomolar range. This approach is appealing for concentration-limited biomolecular systems and medium-to-high-affinity binding studies, demonstrated here at 50 nanomolar protein concentration.

AB - Protein interaction studies often require very low concentrations and highly sensitive biophysical methods. Here, we demonstrate that pulse dipolar electron paramagnetic resonance spectroscopy allows measuring dissociation constants in the nanomolar range. This approach is appealing for concentration-limited biomolecular systems and medium-to-high-affinity binding studies, demonstrated here at 50 nanomolar protein concentration.

U2 - 10.1039/D2CC02360A

DO - 10.1039/D2CC02360A

M3 - Article

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VL - 58

SP - 8790

EP - 8793

JO - Chemical Communications

JF - Chemical Communications

IS - 63

ER -

Pulse dipolar EPR for determining nanomolar binding affinities

Abstract

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Projects

ISSF3 - Bela Bode: ISSF3 - Bela Bode

Enabling Shaped Pulse Capability: Enabling Shaped Pulse Capability for Superior Biological Structural Determination Using EPR Spectroscopy.

Understanding sensitivity gains in pulse: Understanding sensitivity gains in pulse EPR on multimeric membrane proteins

Datasets

Pulse dipolar EPR for determining nanomolar binding affinities (dataset)

Research output

Correction to "Pulse dipolar EPR for determining nanomolar binding affinities": (Chemical Communications (2022), 58:63, DOI:10.1039/d2cc02360a)

Cite this