TY - JOUR
T1 - Benchmark Test and Guidelines for DEER/PELDOR Experiments on Nitroxide-Labeled Biomolecules
AU - Schiemann, Olav
AU - Heubach, Caspar A.
AU - Abdullin, Dinar
AU - Ackermann, Katrin
AU - Azarkh, Mykhailo
AU - Bagryanskaya, Elena G.
AU - Drescher, Malte
AU - Endeward, Burkhard
AU - Freed, Jack H.
AU - Galazzo, Laura
AU - Goldfarb, Daniella
AU - Hett, Tobias
AU - Esteban Hofer, Laura
AU - Fábregas Ibáñez, Luis
AU - Hustedt, Eric J.
AU - Kucher, Svetlana
AU - Kuprov, Ilya
AU - Lovett, Janet Eleanor
AU - Meyer, Andreas
AU - Ruthstein, Sharon
AU - Saxena, Sunil
AU - Stoll, Stefan
AU - Timmel, Christiane R.
AU - Di Valentin, Marilena
AU - McHaourab, Hassane S.
AU - Prisner, Thomas F.
AU - Bode, Bela Ernest
AU - Bordignon, Enrica
AU - Bennati, Marina
AU - Jeschke, Gunnar
N1 - Publisher Copyright:
© 2021 American Chemical Society.
PY - 2021/11/3
Y1 - 2021/11/3
N2 - 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.
AB - 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.
UR - https://www.scopus.com/pages/publications/85118623024
U2 - 10.1021/jacs.1c07371
DO - 10.1021/jacs.1c07371
M3 - Article
C2 - 34664948
AN - SCOPUS:85118623024
SN - 0002-7863
VL - 143
SP - 17875
EP - 17890
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 43
ER -