<<< Full experiment listing


PXD011853 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleHuman dihydrofolate reductase is a substrate of protein kinase CK2α
DescriptionDihydrofolate reductase (DHFR) is a prominent molecular target in antitumor, antibacterial and antiprotozoan chemotherapies. Our in silico amino acid sequence and 3D structure analyses revealed the presence of several putative CK2 phosphorylation sites. Indeed, CK2α subunit phosphorylated DHFR in vitro. In order to identify phosphorylation site we used site-directed mutagenesis to obtain several DHFR mutants with predicted CK2-phosphorylable serine or threonine residues substituted with alanines. All enzyme forms were subjected to in vitro phosphorylation by CK2α subunit. The results pointed to serine 168. Mass spectrometry analyses revealed the presence of additional phosphoserine 145. Phosphorylation by CK2α of S145A mutant and lack of phosphorylation of S145A/S168A double mutant may indicate that S145 phosphorylation may occur only when serine 168 is already phosphorylated. The effect of these and other mutations on enzyme catalytic activity was also investigated.
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterDominik Cysewski
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListphosphorylated residue
InstrumentLTQ Orbitrap Velos; Q Exactive
Dataset History
RevisionDatetimeStatusChangeLog Entry
02018-11-27 08:12:14ID requested
12019-04-15 06:36:52announced
Publication List
Skierka K, Wilamowski P, Wielechowska M, Cysewski D, Senkara E, Wińska P, Bretner M, Cieśla J, Human dihydrofolate reductase is a substrate of protein kinase CK2α. Biochem Biophys Res Commun, 513(2):368-373(2019) [pubmed]
Keyword List
submitter keyword: : dihydrofolate reductase, CK2, protein phosphorylation, protein mutants, phosphoproteomics,LC-MS/MS
Contact List
Joanna Cieśla
contact affiliationWarsaw University of Technology, Faculty of Chemistry, 00-664 Warszawa, Noakowskiego 3, Poland
contact emailjciesla@ch.pw.edu.pl
lab head
Dominik Cysewski
contact affiliationInstitute of Biochemistry and Biophysics Polish Academy of Sciences
contact emaildominikcysewski@gmail.com
dataset submitter
Full Dataset Link List
Dataset FTP location
PRIDE project URI
Repository Record List
[ + ]