Cross-validation of distance measurements in proteins by PELDOR/DEER and single-molecule FRET.

ABSTRACT: Pulsed electron-electron double resonance spectroscopy (PELDOR/DEER) and single-molecule Förster resonance energy transfer spectroscopy (smFRET) are frequently used to determine conformational changes, structural heterogeneity, and inter probe distances in biological macromolecules. They provide qualitative information that facilitates mechanistic understanding of biochemical processes and quantitative data for structural modelling. To provide a comprehensive comparison of the accuracy of PELDOR/DEER and smFRET, we use a library of double cysteine variants of four proteins that undergo large-scale conformational changes upon ligand binding. With either method, we use established standard experimental protocols and data analysis routines to determine inter-probe distances in the presence an

SUBMITTER: Peter MF 

PROVIDER: S-EPMC9338047 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature