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NAD+–nicotinamide exchange assays with Hst2 mutants. Recombinant wild-type and mutant Hst2 proteins were assayed by NAD+–nicotinamide exchange assays. The assays were done by incubating 1.3 ng of enzyme, NAD+, radioactive nicotinamide, and chicken histones (substrate, with or without) at 30° for 1 hr. The Q115A, H135A, and E128A mutants show reduced exchange activity while the R45A mutant shows enhanced activity.
