A stable association with PME‐1 may be dispensable for PP2A demethylation – implications for the detection of PP2A methylation and immunoprecipitation

Reversible methyl‐esterification (methylation) of Leu309 in the protein phosphatase 2A catalytic subunit (PP2Ac) is essential for proper biogenesis of the PP2A holoenzyme. Accumulating evidence links PP2Ac methylation to diseases, including cancer and neurodegenerative disorders. Protein phosphatase...

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Bibliographic Details
Published in:FEBS open bio 2018-09, Vol.8 (9), p.1486-1496
Main Authors: Yabe, Ryotaro, Tsuji, Shunya, Mochida, Satoru, Ikehara, Tsuyoshi, Usui, Tatsuya, Ohama, Takashi, Sato, Koichi
Format: Article
Language:English
Subjects:
Alzheimer's disease
Animal care
Antibiotics
Antigens
Demethylation
Enzymes
Esterase
Esterification
Immunoglobulins
Immunoprecipitation
Methylation
Neurodegenerative diseases
Phosphatase
Phosphoprotein phosphatase
Plasmids
protein methylation
Protein phosphatase
protein phosphatase 2A
protein phosphatase methyl‐esterase
Proteins
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Summary:Reversible methyl‐esterification (methylation) of Leu309 in the protein phosphatase 2A catalytic subunit (PP2Ac) is essential for proper biogenesis of the PP2A holoenzyme. Accumulating evidence links PP2Ac methylation to diseases, including cancer and neurodegenerative disorders. Protein phosphatase methyl‐esterase (PME‐1) specifically catalyzes PP2Ac demethylation. We demonstrate that PP2Ac is demethylated in cell extracts even at 0 °C unless prevented by a PME‐1 methyl‐esterase inhibitor. This promotes dissociation of PP2A heterotrimers with B55 or PR72 subunits, but not those with B56 subunits. These results reveal differential sensitivity of ABC heterotrimers to methylation status of the C subunit. Our study advocates caution when interpreting earlier findings, offers an effective protocol for preserving PP2A complexes, and reveals key distinctions between B subunits and their interactions with the AC core dimer of PP2A. Deregulation of the protein methylation of PP2A is linked to diseases, including cancer and neurodegenerative disorders, necessitating an assay to precisely determine PP2A methylation levels. Here, we show that addition of PME‐1 methyl‐esterase inhibitor ABL127 is necessary for the detection of PP2Ac methylation and immunoprecipitation in vitro; however, blocking the stable PME‐1/PP2A association by PP2A inhibitor okadaic acid is not enough.
ISSN:2211-5463
2211-5463
DOI:10.1002/2211-5463.12485