PXD016409-1

PXD016409 is an original dataset announced via ProteomeXchange.

Title	An identification of kinases which phosphorylates Ebola virus VP30 protein
Description	Samples for mass spectrometry were prepared as follows. HEK293 cells (150 cm^2 dish) were transfected with 6 μg of each protein-coding plasmid by using TransIT (Mirus). Cell lysis and elution were performed as described above. For loading the samples onto an SDS-PAGE, glycerol was added to the eluted supernatants and PAGE was started. Immediately after samples had entered the separation gel, electrophoresis was stopped. After staining the gel with colloidal coomassie, the protein bands were excised and subjected to in-gel digestion with trypsin. Mass spectrometric analysis was performed as described using an Orbitrap Velos Pro mass spectrometer (Thermo Fisher Scientific) which was connected online with a nano C18 RP column equipped Ultimate nanoRSLC-HPLC (Rapid separation liquid chromatography-high performance liquid chromatography) system (Dionex). An aliquot of 15 µL of the tryptic digest was usually injected onto a C18 pre-concentration column and automated trapping and desalting of the sample was performed at a flowrate of 6 µL/min using water/0.05% formic acid as solvent. Tryptic peptides were separated with water/0.045% formic acid and 80% acetonitrile/0.05% formic acid at a flow rate of 300 nL/min. The column was connected to a stainless steel nanoemitter (Proxeon, Denmark) and the eluent sprayed directly towards the heated capillary of the mass spectrometer using a potential of 2300 V. A survey scan with a resolution of 60,000 within the Orbitrap mass analyzer was combined with at least three data-dependent MS/MS scans with dynamic exclusion for 30 s either using CID (Collision-induced dissociation) with the linear ion-trap or using HCD (Higher energy collisional dissociation) and orbitrap detection at a resolution of 7,500. Data analysis was performed using Proteome Discoverer (v4.0; Thermo Fisher Scientific) with SEQUEST and MASCOT (v2.4; Matrix science) search engines using either SwissProt or NCBI databases.
HostingRepository	jPOST
AnnounceDate	2020-02-03
AnnouncementXML	Submission_2020-02-02_23:24:37.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Yuki
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	S-carboxamidomethyl-L-cysteine; O-phospho-L-serine
Instrument	LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2019-11-22 10:41:04	ID requested
⏵ 1	2020-02-02 23:24:38	announced
2	2021-02-04 17:24:33	announced	2021-02-04: Updated FTP location.
3	2022-09-18 03:43:39	announced	2022-09-18: Updated FTP location.

Dataset with its publication pending

submitter keyword: Ebola virus, VP30, immunoprecipitation, phosphorylation

Stephan Becker
lab head
Yuki
contact affiliation	Philipps University Marburg
dataset submitter

jPOST dataset URI

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://jpost.pharm.kyoto-u.ac.jp/repository/JPST000700