Abstract
Publisher Summary This chapter describes methods (light absorption and electron paramagnetic resonance (EPR) spectroscopy) for monitoring the various types of tyrosyl radicals in class I RNRs. These radicals are important targets for antiproliferative compounds. Three types of tyrosyl radicals have been observed among a large number of purified reductases. R2 proteins from E. coli, Mycobacterium tuberculosis, and Arabidopsis thaliana may be considered as representatives of each of these classes. The three proteins can be prepared from overexpressing E. coli strains, which have been transformed with the corresponding plasmids: pVNR2,14 pMtbR2,11,12, and pETR2,13 respectively. The presence of a stable tyrosyl radical in class I RNR can be easily concluded from its X-band EPR spectrum at liquid helium temperature, which shows a characteristic doublet centered at g = 2.0. The experimental conditions for recording the EPR spectrum of the various radicals are discussed in the chapter.
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