Cryoradiolytic reduction of heme proteins: Maximizing dose-dependent yield

Radiolytic reduction in frozen solutions and crystals is a useful method for generation of trapped intermediates in protein-based radical reactions. In this communication we define the conditions which provide the maximum yield of one electron-reduced myoglobin at 77 K using 60Co γ-irradiation in aqueous glycerol glass. The yield reached 50% after 20 kGy, was almost complete at ∼160 kGy total dose, and does not depend on the protein concentration in the range 0.01–5 mM.