DEER sensitivity between iron centers and nitroxides in heme-containing proteins improves dramatically using broadband, high-field EPR

Claire Louise Motion; Janet Eleanor Lovett; Stacey Bell; Scott Lindsay Cassidy; Paul Alexander Sawchuk Cruickshank; David Robert Bolton; Robert Iain Hunter; Hassane El Mkami; Sabine Van Doorslaer; Graham Murray Smith

doi:10.1021/acs.jpclett.6b00456

DEER sensitivity between iron centers and nitroxides in heme-containing proteins improves dramatically using broadband, high-field EPR

Claire Louise Motion, Janet Eleanor Lovett, Stacey Bell, Scott Lindsay Cassidy, Paul Alexander Sawchuk Cruickshank, David Robert Bolton, Robert Iain Hunter, Hassane El Mkami, Sabine Van Doorslaer, Graham Murray Smith

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Abstract

This work demonstrates the feasibility of making sensitive nanometer distancemeasurements between Fe(III) heme centers and nitroxide spin labels in proteins using the double electron-electron resonance (DEER) pulsed EPR technique at 94 GHz. Techniques to measure accurately long distances in many classes of heme proteins using DEER are currently strongly limited by sensitivity. In this paper we demonstrate sensitivity gains of more than 30times compared with previous lower frequency (X-band) DEER measurements on both human neuroglobin and sperm whale myoglobin. This is achieved by: taking advantage of recent instrumental advances; employing wideband excitation techniques based on composite pulses and exploiting more favorable relaxation properties of low-spin Fe(III) in high magnetic fields. This gain in sensitivity potentially allows the DEER technique to be routinely used as a sensitive probe of structure and conformation in the large number of heme and many other metalloproteins.

Original language	English
Pages (from-to)	1411-1415
Number of pages	5
Journal	Journal of Physical Chemistry Letters
Volume	7
Issue number	8
Early online date	1 Apr 2016
DOIs	https://doi.org/10.1021/acs.jpclett.6b00456
Publication status	Published - 21 Apr 2016

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Motion_2016_DEERsensitivity_JPCL_CC
Copyright 2016 American Chemical Society. This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
Final published version, 1.13 MBLicence: CC BY

New Strategies for Spin Labelling: New strategies for spin labelling cysteine-rich proteins
Lovett, J. E. (PI)
EPSRC
1/09/14 → 31/08/15
Project: Standard
Royal Society Research Fellowship: Royal Society Research Fellowship
Lovett, J. E. (PI)
The Royal Society
1/05/14 → 15/02/17
Project: Fellowship
Royal Society Research Fellowship: Royal Society University Research Fellowship
Lovett, B. W. (PI)
The Royal Society
1/09/13 → 30/09/14
Project: Fellowship

Janet Eleanor Lovett
- jel20st-andrews.acuk
- School of Physics and Astronomy - Senior Lecturer
- Biomedical Sciences Research Complex
Person: Academic, Academic - Research
Graham Murray Smith
- gmsst-andrews.acuk
- School of Physics and Astronomy - Professor
Person: Academic

Data underpinning- DEER sensitivity between Iron Centers and Nitroxides in Heme-Containing Proteins Improves Dramatically Using Broadband, High-Field EPR
Motion, C. L. (Creator), Lovett, J. E. (Creator), Bell, S. (Creator), Cassidy, S. L. (Contributor), Cruickshank, P. A. S. (Contributor), Bolton, D. R. (Contributor), Hunter, R. I. (Contributor), El Mkami, H. (Contributor), Van Doorslaer, S. (Contributor) & Smith, G. M. (Contributor), University of St Andrews, 25 Apr 2016
DOI: 10.17630/f6ea167c-132a-4a4d-af00-d2ab30d7cdeb
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Motion, C. L., Lovett, J. E., Bell, S., Cassidy, S. L., Cruickshank, P. A. S., Bolton, D. R., Hunter, R. I., El Mkami, H., Van Doorslaer, S., & Smith, G. M. (2016). DEER sensitivity between iron centers and nitroxides in heme-containing proteins improves dramatically using broadband, high-field EPR. Journal of Physical Chemistry Letters, 7(8), 1411-1415. https://doi.org/10.1021/acs.jpclett.6b00456

@article{9edcefd242874822a219d87594cc1154,

title = "DEER sensitivity between iron centers and nitroxides in heme-containing proteins improves dramatically using broadband, high-field EPR",

abstract = "This work demonstrates the feasibility of making sensitive nanometer distancemeasurements between Fe(III) heme centers and nitroxide spin labels in proteins using the double electron-electron resonance (DEER) pulsed EPR technique at 94 GHz. Techniques to measure accurately long distances in many classes of heme proteins using DEER are currently strongly limited by sensitivity. In this paper we demonstrate sensitivity gains of more than 30times compared with previous lower frequency (X-band) DEER measurements on both human neuroglobin and sperm whale myoglobin. This is achieved by: taking advantage of recent instrumental advances; employing wideband excitation techniques based on composite pulses and exploiting more favorable relaxation properties of low-spin Fe(III) in high magnetic fields. This gain in sensitivity potentially allows the DEER technique to be routinely used as a sensitive probe of structure and conformation in the large number of heme and many other metalloproteins.",

author = "Motion, {Claire Louise} and Lovett, {Janet Eleanor} and Stacey Bell and Cassidy, {Scott Lindsay} and Cruickshank, {Paul Alexander Sawchuk} and Bolton, {David Robert} and Hunter, {Robert Iain} and {El Mkami}, Hassane and {Van Doorslaer}, Sabine and Smith, {Graham Murray}",

note = "Claire Motion would like to acknowledge funding from EPSRC as part of the iMR-CDT. Stacey Bell and Janet Lovett thank EPSRC Grant Number EP/LO22044/1. Janet Lovett also thanks the Royal Society for a University Research Fellowship and Research Grant RG120645. Sabine Van Doorslaer acknowledges the Research Foundation – Flanders (FWO) for financial support (grant G.0687.13). The W-band instrument was developed under the UK Research Council{\textquoteright}s Basic Technology Program (grant EP/F039034/1). We also thank the Wellcome Trust (grant 099149/Z/12/Z). ",

year = "2016",

month = apr,

day = "21",

doi = "10.1021/acs.jpclett.6b00456",

language = "English",

volume = "7",

pages = "1411--1415",

journal = "Journal of Physical Chemistry Letters",

issn = "1948-7185",

publisher = "American Chemical Society (ACS)",

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Motion, CL, Lovett, JE, Bell, S, Cassidy, SL, Cruickshank, PAS, Bolton, DR, Hunter, RI , El Mkami, H, Van Doorslaer, S & Smith, GM 2016, 'DEER sensitivity between iron centers and nitroxides in heme-containing proteins improves dramatically using broadband, high-field EPR', Journal of Physical Chemistry Letters, vol. 7, no. 8, pp. 1411-1415. https://doi.org/10.1021/acs.jpclett.6b00456

TY - JOUR

T1 - DEER sensitivity between iron centers and nitroxides in heme-containing proteins improves dramatically using broadband, high-field EPR

AU - Motion, Claire Louise

AU - Lovett, Janet Eleanor

AU - Bell, Stacey

AU - Cassidy, Scott Lindsay

AU - Cruickshank, Paul Alexander Sawchuk

AU - Bolton, David Robert

AU - Hunter, Robert Iain

AU - El Mkami, Hassane

AU - Van Doorslaer, Sabine

AU - Smith, Graham Murray

N1 - Claire Motion would like to acknowledge funding from EPSRC as part of the iMR-CDT. Stacey Bell and Janet Lovett thank EPSRC Grant Number EP/LO22044/1. Janet Lovett also thanks the Royal Society for a University Research Fellowship and Research Grant RG120645. Sabine Van Doorslaer acknowledges the Research Foundation – Flanders (FWO) for financial support (grant G.0687.13). The W-band instrument was developed under the UK Research Council’s Basic Technology Program (grant EP/F039034/1). We also thank the Wellcome Trust (grant 099149/Z/12/Z).

PY - 2016/4/21

Y1 - 2016/4/21

N2 - This work demonstrates the feasibility of making sensitive nanometer distancemeasurements between Fe(III) heme centers and nitroxide spin labels in proteins using the double electron-electron resonance (DEER) pulsed EPR technique at 94 GHz. Techniques to measure accurately long distances in many classes of heme proteins using DEER are currently strongly limited by sensitivity. In this paper we demonstrate sensitivity gains of more than 30times compared with previous lower frequency (X-band) DEER measurements on both human neuroglobin and sperm whale myoglobin. This is achieved by: taking advantage of recent instrumental advances; employing wideband excitation techniques based on composite pulses and exploiting more favorable relaxation properties of low-spin Fe(III) in high magnetic fields. This gain in sensitivity potentially allows the DEER technique to be routinely used as a sensitive probe of structure and conformation in the large number of heme and many other metalloproteins.

AB - This work demonstrates the feasibility of making sensitive nanometer distancemeasurements between Fe(III) heme centers and nitroxide spin labels in proteins using the double electron-electron resonance (DEER) pulsed EPR technique at 94 GHz. Techniques to measure accurately long distances in many classes of heme proteins using DEER are currently strongly limited by sensitivity. In this paper we demonstrate sensitivity gains of more than 30times compared with previous lower frequency (X-band) DEER measurements on both human neuroglobin and sperm whale myoglobin. This is achieved by: taking advantage of recent instrumental advances; employing wideband excitation techniques based on composite pulses and exploiting more favorable relaxation properties of low-spin Fe(III) in high magnetic fields. This gain in sensitivity potentially allows the DEER technique to be routinely used as a sensitive probe of structure and conformation in the large number of heme and many other metalloproteins.

U2 - 10.1021/acs.jpclett.6b00456

DO - 10.1021/acs.jpclett.6b00456

M3 - Article

SN - 1948-7185

VL - 7

SP - 1411

EP - 1415

JO - Journal of Physical Chemistry Letters

JF - Journal of Physical Chemistry Letters

IS - 8

ER -

DEER sensitivity between iron centers and nitroxides in heme-containing proteins improves dramatically using broadband, high-field EPR

Abstract

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Projects

New Strategies for Spin Labelling: New strategies for spin labelling cysteine-rich proteins

Royal Society Research Fellowship: Royal Society Research Fellowship

Royal Society Research Fellowship: Royal Society University Research Fellowship

Profiles

Janet Eleanor Lovett

Graham Murray Smith

Datasets

Data underpinning- DEER sensitivity between Iron Centers and Nitroxides in Heme-Containing Proteins Improves Dramatically Using Broadband, High-Field EPR

Cite this