Benchmark test and guidelines for DEER/PELDOR experiments on nitroxide-labeled biomolecules
Date
19/10/2021Author
Grant ID
BB/T017740/1
BB/R013780/1
RPG-2018-397
UF090121
Keywords
Metadata
Show full item recordAbstract
Distance distribution information obtained by pulsed dipolar EPR spectroscopy provides an important contribution to many studies in structural biology. Increasingly, such information is used in integrative structural modeling, where it delivers unique restraints on the width of conformational ensembles. In order to ensure reliability of the structural models and of biological conclusions, we herein define quality standards for sample preparation and characterization, for measurements of distributed dipole–dipole couplings between paramagnetic labels, for conversion of the primary time-domain data into distance distributions, for interpreting these distributions, and for reporting results. These guidelines are substantiated by a multi-laboratory benchmark study and by analysis of data sets with known distance distribution ground truth. The study and the guidelines focus on proteins labeled with nitroxides and on double electron–electron resonance (DEER aka PELDOR) measurements and provide suggestions on how to proceed analogously in other cases.
Citation
Schiemann , O , Heubach , C , Abdullin , D , Ackermann , K , Azarkh , M , Bagryanskaya , E , Drescher , M , Endeward , B , Freed , J , Galazzo , L , Goldfarb , D , Hett , T , Esteban Hofer , L , Fábregas Ibáñez , L , Hustedt , E , Kucher , S , Kuprov , I , Lovett , J E , Meyer , A , Ruthstein , S , Saxena , S , Stoll , S , Timmel , C , Di Valentin , M , Mchaourab , H , Prisner , T , Bode , B E , Bordignon , E , Bennati , M & Jeschke , G 2021 , ' Benchmark test and guidelines for DEER/PELDOR experiments on nitroxide-labeled biomolecules ' , Journal of the American Chemical Society , vol. Articles ASAP . https://doi.org/10.1021/jacs.1c07371
Publication
Journal of the American Chemical Society
Status
Peer reviewed
ISSN
0002-7863Type
Journal article
Description
We acknowledge the DFG (German Research Foundation) priority program SPP 1601 for funding multiple projects (O.S., G.J., E.B., M.D., T.F.P. and M.B.) related to the present work together with the funding of many collaborations outside Germany, particularly the NSF (National Science Foundation) funded SharedEPR. JEL thanks The Royal Society for a University Research Fellowship. We thank the Leverhulme Trust (RPG-2018-397) and the BBSRC (BB/R013780/1 and BB/T017740/1) for funding.Collections
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