Orientation-Selective and Frequency-Correlated Light-Induced Pulsed Dipolar Spectroscopy

We explore the potential of orientation-resolved pulsed dipolar spectroscopy (PDS) in light-induced versions of the experiment. The use of triplets as spin-active moieties for PDS offers an attractive tool for studying biochemical systems containing optically active cofactors. Cofactors are often rigidly bound within the protein structure, providing an accurate positional marker. The rigidity leads to orientation selection effects in PDS, which can be analyzed to give both distance and mutual orientation information. Herein we present a comprehensive analysis of the orientation selection of a full set of light-induced PDS experiments. We exploit the complementary information provided by the different light-induced techniques to yield atomic-level structural information. For the first time, we measure a 2D frequency-correlated laser-induced magnetic dipolar spectrum, and we are able to monitor the complete orientation dependence of the system in a single experiment. Alternatively, the summed spectrum enables an orientation-independent analysis to determine the distance distribution.

Automated summary

The study explores the potential of orientation-resolved pulsed dipolar spectroscopy (PDS) in light-induced experiments, using triplets as spin-active moieties for studying biochemical systems. Analysis of orientation selection effects in PDS provides both distance and mutual orientation information, with complementary information from different light-induced techniques yielding atomic-level structural information. The study introduces a 2D frequency-correlated laser-induced magnetic dipolar spectrum, allowing for monitoring of complete orientation dependence in a single experiment, as well as an orientation-independent analysis for determining distance distribution.