PXD011853-1

PXD011853 is an original dataset announced via ProteomeXchange.

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

Revision	Datetime	Status	ChangeLog Entry
0	2018-11-27 08:12:14	ID requested
⏵ 1	2019-04-15 06:36:52	announced

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]

submitter keyword: : dihydrofolate reductase, CK2, protein phosphorylation, protein mutants, phosphoproteomics,LC-MS/MS

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

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/04/PXD011853

PRIDE project URI

• PRIDE
  1. PXD011853
     1. Label: PRIDE project
     2. Name: Human dihydrofolate reductase is a substrate of protein kinase CK2α