PXD011853 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Human dihydrofolate reductase is a substrate of protein kinase CK2α |
Description | Dihydrofolate 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. |
HostingRepository | PRIDE |
AnnounceDate | 2019-04-15 |
AnnouncementXML | Submission_2019-04-15_06:36:51.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Dominik Cysewski |
SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: 9606; |
ModificationList | phosphorylated residue |
Instrument | LTQ Orbitrap Velos; Q Exactive |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2018-11-27 08:12:14 | ID requested | |
⏵ 1 | 2019-04-15 06:36:52 | announced | |
Publication List
Skierka K, Wilamowski P, Wielechowska M, Cysewski D, Senkara E, Wi, ń, ska P, Bretner M, Cie, ś, la J, . 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 affiliation | Warsaw University of Technology, Faculty of Chemistry, 00-664 Warszawa, Noakowskiego 3, Poland |
contact email | jciesla@ch.pw.edu.pl |
lab head | |
Dominik Cysewski |
contact affiliation | Institute of Biochemistry and Biophysics Polish Academy of Sciences |
contact email | dominikcysewski@gmail.com |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD011853
- Label: PRIDE project
- Name: Human dihydrofolate reductase is a substrate of protein kinase CK2α