A Global Minimization Toolkit for Batch-Fitting and χ2 Cluster Analysis of CW-EPR Spectra

Abstract

Electron paramagnetic resonance spectroscopy (EPR) is a uniquely powerful technique for characterizing conformational dynamics at specific sites within a broad range of molecular species in water. Computational tools for fitting EPR spectra have enabled dynamics parameters to be determined quantitatively. These tools have dramatically broadened the capabilities of EPR dynamics analysis, however, their implementation can easily lead to overfitting or problems with self-consistency. As a result, dynamics parameters and associated properties become difficult to reliably determine, particularly in the slow-motion regime. Here, we present an EPR analysis strategy and the corresponding computational tool for batch-fitting EPR spectra and cluster analysis of the χ2 landscape in Linux. We call this tool CSCA (Chi-Squared Cluster Analysis). The CSCA tool allows us to determine self-consistent rotational diffusion rates and enables calculations of activation energies of diffusion from Arrhenius plots. We demonstrate CSCA using a model system designed for EPR analysis: a self-assembled nanoribbon with radical electron spin labels positioned at known distances off the surface. We anticipate that the CSCA tool will increase the reproducibility of EPR fitting for the characterization of dynamics in biomolecules and soft matter.