Pulse electron paramagnetic resonance measurements of nanometer scale distance distributions have proven highly effective in structural studies. They exploit the magnetic dipole-dipole coupling between spin labels site-specifically attached to macromolecules. The most commonly applied technique is double electron-electron resonance (DEER, also called pulsed electron double resonance (PELDOR)). Here we present the new technique of laser-induced magnetic dipole (LaserIMD) spectroscopy based on optical switching of the dipole-dipole coupling. In a proof of concept experiment on a model peptide, we find, already at a low quantum yield of triplet excitation, the same sensitivity for measuring the distance between a porphyrin and a nitroxide label as in a DEER measurement between two nitroxide labels. On the heme protein cytochrome C, we demonstrate that LaserIMD allows for distance measurements between a heme prosthetic group and a nitroxide label, although the heme triplet state is not directly observable by an electron spin echo.
Following the first evidence of the triplet character of the optically excited phosphorescent state of naphthalene by Hutchison and Mangum in 1958, electron paramagnetic resonance (EPR) spectroscopy has become widely used to study and understand the properties and kinetic characteristics of excited triplet states. This minireview gives an overview over EPR techniques based on continuous microwave methods using lock-in or direct detection as well as pulsed EPR methods with respect to their suitability for kinetic studies of excited triplet states. A short historical overview of the experimental and theoretical developments in this field of research as well as of the triplet systems studied, with a final focus on fullerenes, is given. This may help newcomers to the field as a guide to the relevant literature.
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