PXD045961

PXD045961 is an original dataset announced via ProteomeXchange.

Title	Overexpress S-protein tagged mouse TBK1 protein and identify the TBK1-interacting proteins by mass spectrometry
Description	HEK293T cells were transfected with SFB-tagged mTBK1 or an empty vector for 24 h, the cells were lysed with lysis buffer (20 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.5% Triton X-100, 10% glycerol, 1 mM ethylenediaminetetraacetic acid) containing a complete protease inhibitor cocktail, followed by centrifugation at 16000xg for 10 min at 4 °C. The supernatants were subjected to immunoprecipitation using S-protein agarose (Millipore, Cat# 69704) for 3 h at 4 °C. Immunoprecipitates were washed with lysis buffer containing 150 mM NaCl three times and incubated with cell lysates from RAW 264.7 cells overnight at 4 °C. The immunoprecipitated complexes were washed three times again and separated by SDS-PAGE, and the gel was stained with Coomassie brilliant blue. The entire lane was cut into 2-mm gel slices, digested with Trypsin, and subjected to LC-MS/MS assays using an Orbitrap Elite mass spectrometer (Thermo Fisher Scientific). The mass spectrometry data were analyzed using Thermo Proteome Discovery (Version2.3), and tandem mass spectra were searched against the UniProt- the UniProt Mus musculus database 2022.11.16.
HostingRepository	iProX
AnnounceDate	2023-10-27
AnnouncementXML	Submission_2023-12-04_20:58:47.283.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jing Yang
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606; scientific name: Mus musculus; NCBI TaxID: 10090;
ModificationList	No PTMs are included in the dataset
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2023-10-07 20:16:21	ID requested
1	2023-10-07 20:16:39	announced
⏵ 2	2023-12-04 20:58:48	announced	2023-12-05: Update publication information.

Lin Y, Yang J, Yang Q, Zeng S, Zhang J, Zhu Y, Tong Y, Li L, Tan W, Chen D, Sun Q, PTK2B promotes TBK1 and STING oligomerization and enhances the STING-TBK1 signaling. Nat Commun, 14(1):7567(2023) [pubmed]

submitter keyword: SFB-tagged mTBK1, RAW 264.7 cells, LC-MS/MS assays

Qinmiao Sun
contact affiliation	Institute of Zoology, Chinese Academy of Sciences
contact email	qinmiaosun@ioz.ac.cn
lab head
Jing Yang
contact affiliation	Institute of Zoology, Chinese Academy of Sciences
contact email	yangjing22@ioz.ac.cn
dataset submitter

iProX dataset URI