Physics

Nanotechnology

Dipole

Site-directed spin labeling

Organic chemistry

Biological system

Characterization (materials science)

Biomolecule

Biology

Geodesy

Radical polymerization

Benchmark (surveying)

Copolymer

Chemical physics

Power (physics)

Geography

Nitroxide mediated radical polymerization

Chemistry

Electron paramagnetic resonance

Materials science

Nuclear magnetic resonance

Reliability (semiconductor)

Polymer

Biochemistry

Thermodynamics

Journal of the American Chemical Society

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.

Benchmark Test and Guidelines for DEER/PELDOR Experiments on Nitroxide-Labeled Biomolecules