Increasing the Modulation Depth of GdIII-Based Pulsed Dipolar EPR Spectroscopy (PDS) with Porphyrin-GdIII Laser-Induced Magnetic Dipole Spectroscopy

Abstract

Distance determination with pulsed EPR has become an important technique for the structural investigation of biomacromolecules, with double electron-electron resonance spectroscopy (DEER) as the most important method. Gd^III-based spin labels are one of the most frequently used spin labels for DEER owing to their stability against reduction, high magnetic moment, and absence of orientation selection. A disadvantage of Gd^III-Gd^III DEER is the low modulation depth due to the broad EPR spectrum of Gd^III. Here, we introduce laser-induced magnetic dipole spectroscopy (LaserIMD) with a spin pair consisting of Gd^III(PymiMTA) and a photoexcited porphyrin as an alternative technique. We show that the excited state of the porphyrin is not disturbed by the presence of the Gd^III complex and that herewith modulation depths of almost 40% are possible. This is significantly higher than the value of 7.2% that was achieved with Gd^III-Gd^III DEER.