PP2AC Phospho-Tyr307 Antibodies Are Not Specific for this Modification but Are Sensitive to Other PP2AC Modifications Including Leu309 Methylation | Zendy

Ingrid E. Frohner | Zendy; Ingrid Mudrak | Zendy; Stefan Schüchner | Zendy; Dorothea Anrather | Zendy; Markus Hartl | Zendy; JeanMarie Sontag | Zendy; Estelle Sontag | Zendy; Brian E. Wadzinski | Zendy; Teresa Preglej | Zendy; Wilfried Ellmeier | Zendy; Egon Ogris | Zendy

Open Access

PP2AC Phospho-Tyr307 Antibodies Are Not Specific for this Modification but Are Sensitive to Other PP2AC Modifications Including Leu309 Methylation

Author(s) -

Ingrid E. Frohner,

Ingrid Mudrak,

Stefan Schüchner,

Dorothea Anrather,

Markus Hartl,

JeanMarie Sontag,

Estelle Sontag,

Brian E. Wadzinski,

Teresa Preglej,

Wilfried Ellmeier,

Egon Ogris

Publication year - 2020

Publication title -

cell reports

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 6.264

H-Index - 154

eISSN - 2639-1856

pISSN - 2211-1247

DOI - 10.1016/j.celrep.2020.02.035

Subject(s) - protein phosphatase 2 , phosphorylation , phosphatase , protein subunit , monoclonal antibody , antibody , immunoprecipitation , biochemistry , chemistry , threonine , protein phosphorylation , biology , protein tyrosine phosphatase , methylation , microbiology and biotechnology , protein kinase a , serine , immunology , gene

Protein phosphatase 2A (PP2A) is an important regulator of signal transduction pathways and a tumor suppressor. Phosphorylation of the PP2A catalytic subunit (PP2A C ) at tyrosine 307 has been claimed to inactivate PP2A and was examined in more than 180 studies using commercial antibodies, but this modification was never identified using mass spectrometry. Here we show that the most cited pTyr 307 monoclonal antibodies, E155 and F-8, are not specific for phosphorylated Tyr 307 but instead are hampered by PP2A C methylation at leucine 309 or phosphorylation at threonine 304. Other pTyr 307 antibodies are sensitive to PP2A C methylation as well, and some cross-react with pTyr residues in general, including phosphorylated hemagglutinin tags. We identify pTyr 307 using targeted mass spectrometry after transient overexpression of PP2A C and Src kinase. Yet under such conditions, none of the tested antibodies show exclusive pTyr 307 specificity. Thus, data generated using these antibodies need to be revisited, and the mechanism of PP2A inactivation needs to be redefined.

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