Abstract

Here, we introduce FRETpredict, a Python software program to predict FRET efficiencies from ensembles of protein conformations. FRETpredict uses an established Rotamer Library Approach to describe the FRET probes covalently bound to the protein. The software efficiently operates on large conformational ensembles such as those generated by molecular dynamics simulations to facilitate the validation or refinement of molecular models and the interpretation of experimental data. We demonstrate the performance and accuracy of the software for different types of systems: a relatively structured peptide (polyproline 11), an intrinsically disordered protein (ACTR), and three folded proteins (HiSiaP, SBD2, and MalE). We also describe a general approach to generate new rotamer libraries for FRET probes of interest. FRETpredict is open source (GPLv3) and is available at github.com/KULL-Centre/FRETpredict and as a Python PyPI package at pypi.org/project/FRETpredict. Author SummaryWe present FRETpredict, an open-source software to calculate FRET observables from protein structures. Using a previously developed Rotamer Library Approach, FRETpredict helps place multiple conformations of the selected FRET probes at the labeled sites, and use these to calculate FRET efficiencies. Through several case studies, we illustrate the ability of FRETpredict to interpret experimental results and validate protein conformations. We also explain a methodology for generating new rotamer libraries of FRET probes of interest.