Benchmark Test and Guidelines for DEER/PELDOR Experiments on Nitroxide-Labeled Biomolecules
Site-directed spin labeling
Benchmark (surveying)
Biomolecule
Characterization
DOI:
10.1021/jacs.1c07371
Publication Date:
2021-10-21T00:15:19Z
AUTHORS (30)
ABSTRACT
Distance distribution information obtained by pulsed dipolar EPR spectroscopy provides an important contribution to many studies in structural biology. Increasingly, such is used integrative modeling, where it delivers unique restraints on the width of conformational ensembles. In order ensure reliability models and biological conclusions, we herein define quality standards for sample preparation characterization, measurements distributed dipole-dipole couplings between paramagnetic labels, conversion primary time-domain data into distance distributions, interpreting these reporting results. These guidelines are substantiated a multi-laboratory benchmark study analysis sets with known ground truth. The focus proteins labeled nitroxides double electron-electron resonance (DEER aka PELDOR) provide suggestions how proceed analogously other cases.
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