PXD014879

PXD014879 is an original dataset announced via ProteomeXchange.

Title	Characterization of the C-terminal post-translational modification pattern of the catalytic C subunit of protein phosphatase 2A by mass spectrometry.
Description	Protein phosphatase 2A (PP2A) is an important regulator of cellular signal transduction pathways. The carboxy-terminal tail of the PP2A catalytic C subunit is thought to be post-translationally modified at three critical sites. Carboxy-terminal methyl-esterification at Leucine 309 is essential for the assembly of most trimeric PP2A complexes. Two phosphorylation sites have been identified at threonine 304 and tyrosine 307. Phosphorylation at Tyr307 has been claimed to inactivate PP2A and has been examined in over 180 studies using commercial antibodies. Yet, surprisingly, Tyr307 phosphorylation has never been identified by mass spectrometry analysis. This fact raised our concern about the existence of this modification in vivo and also about the specificity of antibodies used to detect this modification. For the first time we were able to identify by mass spectrometry phosphorylation of Tyr307 upon transient overexpression of active Src kinase and detected also tyrosine phosphorylation in hemagglutinin (HA) tags. In addition we were able to identify carboxy-terminal methyl-esterification at leucine 309 and also confirmed phosphorylation at threonine 304. Our analysis of commercial antibodies against pTyr307 revealed that none of the tested antibodies shows the designated specificity solely for pTyr307 and thus all data generated by these antibodies need to be revisited.
HostingRepository	PRIDE
AnnounceDate	2020-03-03
AnnouncementXML	Submission_2020-03-24_00:40:52.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Dorothea Anrather
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	phosphorylated residue; L-leucine methyl ester
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2019-08-01 07:28:53	ID requested
1	2020-03-02 23:17:14	announced
⏵ 2	2020-03-24 00:40:53	announced	2020-03-24: Updated publication reference for PubMed record(s): 32130916.

Frohner IE, Mudrak I, Sch, ü, chner S, Anrather D, Hartl M, Sontag JM, Sontag E, Wadzinski BE, Preglej T, Ellmeier W, Ogris E, Methylation. Cell Rep, 30(9):3171-3182.e6(2020) [pubmed]

submitter keyword: Protein phosphatase 2A, tyrosine phosphorylation, pTyr307- specific antibodies, C-terminal protein methylation

Egon Ogris
contact affiliation	Center for Medical Biochemistry, Max Perutz Labs, Medical University of Vienna, Austria
contact email	egon.ogris@univie.ac.at
lab head
Dorothea Anrather
contact affiliation	Max Perutz Laoratories Mass spectrometry facility
contact email	dorothea.anrather@univie.ac.at
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD014879
     1. Label: PRIDE project
     2. Name: Characterization of the C-terminal post-translational modification pattern of the catalytic C subunit of protein phosphatase 2A by mass spectrometry.