PXD039663

PXD039663 is an original dataset announced via ProteomeXchange.

Title	Phosphosite mapping on PI3Kc1 by ULK1 and TBK1 kinases.
Description	Autophagy is a highly conserved catabolic process which degrades and recycles intracellular components. To accomplish this task the surplus material (cargo) is sequestered in a double membranous vesicle termed autophagosome. To initiate the formation of autophagosome autophagy machinery is recruited to cargo in a controlled manner. One of the first factors recruited to damaged mitochondria during mitophagy (selective removal of damaged mitochondria by autophagy) are two protein kinases, ULK1/2 complex and TBK1. They both promote a subsequent recruitment of the class III phosphatidylinositol 3-kinase complex I (PI3Kc1 complex) which function is to generate the lipid phosphatidylinositol 3-phosphate (PI(3)P) on phagophore membranes. Both ULK1 and TBK1 were reported to phosphorylate several autophagy factors. Here, we probed phosphorylation of PI3Kc1 by ULK1 and TBK1 to gain a full picture of the phosphosites map of PI3Kc phosphorylated by both protein kinases. Recombinant PI3Kc1 was incubated with recombinant ULK1 complex or TBK1 in the presence of ATP and MgCl2. As a negative control, the same protein mixtures were incubated in a kinase buffer lacking MgCl2 and ATP. Samples were separated by SDS-PAGE and stained with Coomassie. The bands for each subunit of the PI3KC3-C1 complex (VPS15, VPS34, ATG14, and Beclin1) were extracted from the gel and submitted for mass spectrometry analysis.
HostingRepository	PRIDE
AnnounceDate	2023-05-08
AnnouncementXML	Submission_2023-05-08_03:42:54.318.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	WeiQiangChen
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	Q Exactive HF-X

Revision	Datetime	Status	ChangeLog Entry
0	2023-01-25 08:16:33	ID requested
⏵ 1	2023-05-08 03:42:54	announced

Dataset with its publication pending

submitter keyword: PI3Kc3-C1, phosphorylation,ULK1, TBK1

SaschaMartens
contact affiliation	Max Perutz Labs, Vienna BioCenter, Dr. Bohr-Gasse 9, Vienna, Austria
contact email	sascha.martens@univie.ac.at
lab head
WeiQiangChen
contact affiliation	Mass Spectrometry Facility, Max Perutz Laboratories Support GmbH
contact email	chenweiqiang@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD039663
     1. Label: PRIDE project
     2. Name: Phosphosite mapping on PI3Kc1 by ULK1 and TBK1 kinases.