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  Benchmark test and guidelines for DEER/PELDOR experiments on nitroxide-labeled biomolecules

Schiemann, O., Heubach, C. A., Abdullin, D., Ackermann, K., Azarkh, M., Bagryanskaya, E. G., et al. (2021). Benchmark test and guidelines for DEER/PELDOR experiments on nitroxide-labeled biomolecules. Journal of the American Chemical Society, 143(43), 17875-17890. doi:10.1021/jacs.1c07371.

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 Creators:
Schiemann, O., Author
Heubach, C. A., Author
Abdullin, D., Author
Ackermann, K., Author
Azarkh, M., Author
Bagryanskaya, E. G., Author
Drescher, M., Author
Endeward, B., Author
Freed, J. H., Author
Galazzo, L., Author
Goldfarb, D., Author
Hett, T., Author
Hofer, L. E., Author
Ibáñez, L. F., Author
Hustedt, E. J., Author
Kucher, S., Author
Kuprov, I., Author
Lovett, J. E., Author
Meyer, A.1, Author              
Ruthstein, S., Author
Saxena, S., AuthorStoll, S., AuthorTimmel, C. R., AuthorValentin, M. D., AuthorMchaourab, H. S., AuthorPrisner, T. F., AuthorBode, B. E., AuthorBordignon, E., AuthorBennati, M.1, Author              Jeschke, G., Author more..
Affiliations:
1Research Group of Electron Paramagnetic Resonance, MPI for Biophysical Chemistry, Max Planck Society, ou_578606              

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 Abstract: 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.

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Language(s): eng - English
 Dates: 2021-10-192021-11-03
 Publication Status: Published in print
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 Rev. Type: Peer
 Identifiers: DOI: 10.1021/jacs.1c07371
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Title: Journal of the American Chemical Society
Source Genre: Journal
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Pages: - Volume / Issue: 143 (43) Sequence Number: - Start / End Page: 17875 - 17890 Identifier: -