E275 and F276 in β12-β13 Loop of Protein Phosphatase-1 Resist Mn²⁺-Mediated Activation

Abstract

Protein phosphatase-1 (PP1) is one of the most important Ser/Thr phosphatases in eukaryotic cells. G274, E275, and F276 are located at the tip of the β12-β13 loop of protein phosphatase-1. Without Mn²⁺, the basal activities and intrinsic fluorescence spectra of all single and double deletion mutants of G274, E275, and F276 were similar to those of PP1, but deletion mutants ΔE275 and ΔE275F276 showed hyperactivity and corresponding changes in intrinsic fluorescence spectra when Mn²⁺ was present. This suggests that the conformation transition resulting from the combined effect of mutation and Mn²⁺ accounts for the hyperactivity of mutants. These observations imply that E275 and F276 play a role in resisting enzyme activation by Mn²⁺ in PP1.