Substrate-dependent metal preference of PPM1H, a cancer-associated protein phosphatase 2C: comparison with other family members.
Protein phosphatase 2C (PP2C) family is characterized by requirement of metal cation for phosphatase activity. We previously established that PPM1H is a cancer-associated member of the PP2C family. Here we further characterized the phosphatase activity of PPM1H, focusing on its dependence on metal cation. PPM1H possesses the potential to dephosphorylate p-nitrophenyl phosphate (pNPP), casein and phosphopeptides. Interestingly, PPM1H shows the metal preference that is varied depending on the substrate (substrate-dependent metal preference); PPM1H prefers Mn(2+) when pNPP or phosphopeptides is used as a substrate. Meanwhile, a preference for Mg(2+) is displayed by PPM1H with casein as a substrate. When both cations are added to the reaction, the degree of the effect is always closer to that with Mn(2+) alone, irrespective of the substrate. This preponderance of Mn(2+) is explained by its greater affinity for PPM1H than Mg(2+). From the literature the substrate-dependent metal preference appears to be shared by other PP2Cs. According to the crystal structure, a binuclear metal center of PP2C plays an important role for coordinating the substrate and nucleophilic waters in the active site. Therefore, the differences in the size, preferred geometry and coordination requirements between two metals, in relation to the substrate, may be responsible for this intriguing property.
Sugiura
Takeyuki
T
Discovery Research Laboratory, Tokyo R&D Center, Daiichi Pharmaceutical Co., Ltd., Daiichi-Sankyo Group, Tokyo, Japan. sugiura.takeyuki.uu@daiichisankyo.co.jp
Noguchi
Yoshie
Y
D000814:Aniline Compounds / Q000378:metabolism; D002364:Caseins / Q000378:metabolism; D006801:Humans; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D008670:Metals / Q000378:metabolism*; D009943:Organophosphorus Compounds / Q000378:metabolism; D010748:Phosphopeptides / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D000071636:Protein Phosphatase 2C; D013379:Substrate Specificity
D000814:Aniline Compounds; D002364:Caseins; D008670:Metals; D009943:Organophosphorus Compounds; D010748:Phosphopeptides; D008345:Manganese; C068798:4-aminophenylphosphate; C000607456:PPM1A protein, human; C000607457:PPM1B protein, human; C000607460:PPM1G protein, human; C521058:PPM1H protein, human; D010749:Phosphoprotein Phosphatases; D000071636:Protein Phosphatase 2C; D008274:Magnesium