- Title
- PP2Ac Phospho-Tyr307 Antibodies Are Not Specific for this Modification but Are Sensitive to Other PP2Ac Modifications Including Leu309 Methylation
- Creator
- Frohner, Ingrid E.; Mudrak, Ingrid; Ogris, Egon; Schüchner, Stefan; Anrather, Dorothea; Hartl, Markus; Sontag, Jean-Marie; Sontag, Estelle; Wadzinski, Brian E.; Preglej, Teresa; Ellmeier, Wilfried
- Relation
- Cell Reports Vol. 30, Issue 9, p. 3171-3182.e6
- Publisher Link
- http://dx.doi.org/10.1016/j.celrep.2020.02.035
- Publisher
- Cell Press
- Resource Type
- journal article
- Date
- 2020
- Description
- Protein phosphatase 2A (PP2A) is an important regulator of signal transduction pathways and a tumor suppressor. Phosphorylation of the PP2A catalytic subunit (PP2AC) 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 pTyr307 monoclonal antibodies, E155 and F-8, are not specific for phosphorylated Tyr307 but instead are hampered by PP2AC methylation at leucine 309 or phosphorylation at threonine 304. Other pTyr307 antibodies are sensitive to PP2AC methylation as well, and some cross-react with pTyr residues in general, including phosphorylated hemagglutinin tags. We identify pTyr307 using targeted mass spectrometry after transient overexpression of PP2AC and Src kinase. Yet under such conditions, none of the tested antibodies show exclusive pTyr307 specificity. Thus, data generated using these antibodies need to be revisited, and the mechanism of PP2A inactivation needs to be redefined.
- Subject
- abcam monoclonal E155; methyl-PP2A sensitivity; non-specific antibody; PP2A catalytic subunit; PP2A inactivation; PP2A phospho-tyrosine 307
- Identifier
- http://hdl.handle.net/1959.13/1456997
- Identifier
- uon:45285
- Identifier
- ISSN:2211-1247
- Rights
- © 2020 The Author(s). This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
- Language
- eng
- Full Text
- Reviewed
- Hits: 3093
- Visitors: 3171
- Downloads: 82
| Thumbnail | File | Description | Size | Format | |||
|---|---|---|---|---|---|---|---|
| View Details Download | ATTACHMENT02 | Publisher version (open access) | 2 MB | Adobe Acrobat PDF | View Details Download |