PXD042794-1

PXD042794 is an original dataset announced via ProteomeXchange.

Title	Role of IP6K1 in mammalian gut physiology
Description	AGS cells transiently transfected to express either SFB-tagged GFP or SFB-tagged IP6K1 were lysed in NETN buffer (20 mM Tris-HCl, pH 8.0, 100 mM NaCl, 1 mM EDTA, 0.5% Nonidet P-40) containing protease and phosphatase inhibitor cocktails (Merck), at 4 degree C for 60 min. Cell debris were removed by centrifugation at 14000 x g for 10 min, and cell lysates were incubated with streptavidin-sepharose beads (Cytiva Life Sciences) for 1 h at 4 degree C with end-over-end rotation. The bound protein complexes were washed thrice with NETN buffer and then eluted with 2 mg/mL biotin (Merck) for 90 min at 4 degree C. The eluates were incubated with S-protein agarose beads (Novagen) for 1 h at 4 degree C and then washed thrice with NETN buffer. Proteins bound to S-protein agarose beads were boiled in 2x SDS sample buffer for 5 min, loaded on a 12% SDS polyacrylamide gel, allowed to run into the resolving gel up to 1 cm, and visualized by Coomassie Brilliant Blue staining. All proteins in the sample were excised in one gel slice and sent for mass spectrometry analysis to Taplin Biological Mass Spectrometry Facility at Harvard University, USA. Briefly, gel pieces were subjected to in-gel digestion with sequencing-grade trypsin (Promega), and the extracted peptides were resolved by reverse phase HPLC. Eluted peptides were subjected to electrospray ionization (ESI) and then allowed to enter an LTQ Orbitrap Velos Pro ion-trap mass spectrometer (Thermo Fisher Scientific). Peptides were detected, isolated, and fragmented to produce a tandem mass spectrum of specific fragment ions for each peptide. Peptide sequences (and hence protein identity) were determined by matching protein databases from UniProt (https://www.uniprot.org/taxonomy/10090) with the acquired fragmentation pattern by the software program, Sequest (Thermo Fisher Scientific).
HostingRepository	MassIVE
AnnounceDate	2025-09-17
AnnouncementXML	Submission_2025-09-17_01:21:39.783.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Akruti Shah
SpeciesList	scientific name: Homo sapiens; common name: human; NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	LTQ Orbitrap Velos Pro

Revision	Datetime	Status	ChangeLog Entry
0	2023-06-07 07:15:11	ID requested
⏵ 1	2025-09-17 01:21:40	announced

no publication

submitter keyword: IP6K1, InsP7

Rashna Bhandari
contact affiliation	Centre for DNA Fingerprinting and Diagnostics
contact email	rashna@cdfd.org.in
lab head
Akruti Shah
contact affiliation	National Institutes of Health
contact email	akruti.shah26@gmail.com
dataset submitter

MassIVE 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://massive-ftp.ucsd.edu/v05/MSV000092121/