PXD047996

PXD047996 is an original dataset announced via ProteomeXchange.

Title	Autophosphorylation of Tousled-like kinases (TLK1 and TLK2) regulates recruitment to damaged chromatin via PCNA interaction (Tandem Affinity Purification coupled to LC-MS/MS of SFB-TLK1 WT and TLK2 WT)
Description	S-protein-2xFLAG-Streptavidin binding peptide (SFB)-tagged TLK1 or SFB-TLK2 were transfected into 4x 15 cm plates of HEK293T cells. Cells were harvested 24 hours after transfection using NETN buffer (150 mM NaCl, 0.5 mM EDTA, 20 mM Tris-HCl pH 8.0, 0.5% NP-40) supplemented with 2 µg/mL aprotinin (Thermo, AAJ60237MB) and 5 µg/mL pepstatin A (Thermo, PI78432) at 4ºC for 20 minutes. Lysates were centrifuged at 9,000 x g, 4ºC for 20 minutes to yield supernatant as soluble fraction. Soluble fractions were incubated with streptavidin sepharose (200 µl) (GE Healthcare, GE17-5113-01) at 4ºC for 1 hour followed by washing with NETN buffer three times. The protein complexes were eluted with 2 mg/mL biotin at 4ºC for 1 hour. The eluents were then incubated with S-protein agarose (EMD Millipore, 69704-3) overnight at 4ºC, washed three times with NETN buffer, and eluted in 1x Laemmli buffer.
HostingRepository	PRIDE
AnnounceDate	2025-01-02
AnnouncementXML	Submission_2025-01-02_07:21:15.391.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD047996
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Kirk West
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue; acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2023-12-20 04:31:21	ID requested
⏵ 1	2025-01-02 07:21:16	announced

10.1093/nar/gkae1279;

West KL, Nguyen TTN, Tengler KA, Kreiling N, Raney KD, Ghosal G, Leung JW, Autophosphorylation of the Tousled-like kinases TLK1 and TLK2 regulates recruitment to damaged chromatin via PCNA interaction. Nucleic Acids Res, ():(2024) [pubmed]

10.6019/PXD047996;

submitter keyword: Tandem Affinity purification,Tousled-Like Kinases, LC-MS/MS

Justin W. Leung
contact affiliation	Department of Radiation Oncology Univeristy of Texas Health Science Center at San Antonio
contact email	leungj@uthscsa.edu
lab head
Kirk West
contact affiliation	University of Arkansas for Medical Sciences
contact email	kwest@uams.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD047996
     1. Label: PRIDE project
     2. Name: Autophosphorylation of Tousled-like kinases (TLK1 and TLK2) regulates recruitment to damaged chromatin via PCNA interaction (Tandem Affinity Purification coupled to LC-MS/MS of SFB-TLK1 WT and TLK2 WT)