PP2A(C) Phospho-Tyr(307) Antibodies Are Not Specific for this Modification but Are Sensitive to Other PP2A(C) Modifications Including Leu(309) Methylation

Simple item page
dc.contributor.authorFrohner, Ingrid E.
dc.contributor.authorMudrak, Ingrid
dc.contributor.authorSchuechner, Stefan
dc.contributor.authorAnrather, Dorothea
dc.contributor.authorHartl, Markus
dc.contributor.authorSontag, Jean-Marie
dc.contributor.authorSontag, Estelle
dc.contributor.authorWadzinski, Brian E.
dc.contributor.authorPreglej, Teresa
dc.contributor.authorEllmeier, Wilfried
dc.contributor.authorOgris, Egon
dc.date.accessioned2020-11-03T23:17:57Z
dc.date.available2020-11-03T23:17:57Z
dc.date.issued2020-03-03
dc.description.abstractProtein 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.en_US
dc.description.sponsorshipWe thank Dr. David Virshup, Duke-NUS Medical School (Singapore), for critically reading the manuscript and Dr. John Alberta and Dr. Thomas Roberts, Dana-Farber Cancer Institute (DFCI), Harvard Medical School (Boston), for materials. MS analyses were performed at the Mass Spectrometry Facility of Max Perutz Labs Vienna using the Vienna BioCenter Core Facilities (VBCF) instrument pool. This work was funded by service and royalty fees from antibody licensing agreements and the monoclonal antibody service facility (E.O. lab), and was partially supported by grant G1700055 from the Hunter Medical Research Institute (New South Wales, Australia) (E.S. and J.-M.S.).en_US
dc.identifier.doi10.1016/j.celrep.2020.02.035
dc.identifier.issn2211-1247
dc.identifier.urihttp://hdl.handle.net/1803/16270
dc.language.isoen_USen_US
dc.publisherCell Reportsen_US
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
dc.source.urihttps://www.sciencedirect.com/science/article/pii/S2211124720301960
dc.titlePP2A(C) Phospho-Tyr(307) Antibodies Are Not Specific for this Modification but Are Sensitive to Other PP2A(C) Modifications Including Leu(309) Methylationen_US
dc.typeArticleen_US
Files
Original bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
PP2A(C) Phospho-Tyr(307) Antibodies Are Not Specific for this Modification but Are Sensitive to Other PP2A(C) Modifications Including Leu(309) Methylation.pdf
Size:
2.2 MB
Format:
Adobe Portable Document Format
Description:
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.93 KB
Format:
Item-specific license agreed upon to submission
Description:
Download
Collections
Cell & Developmental Biology