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Addition of PKR enhances protease sensitivity of the Ser51 loop in eIF2α. (A and B) Purified yeast eIF2α3–175 was incubated with thermolysin and the indicated ratio (wt/wt) of purified GST–PKR kinase domain (KD) or mutant GST–PKR–KD–T487D. Protease reaction products were separated by SDS-PAGE and visualized by Coomassie Blue staining. The positions of intact GST–PKR–KD and eIF2α3–175, as well as the eIF2α C-terminal cleavage product (starting at Leu61), are indicated.
