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PXD003496

PXD003496 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleIdentification of the UT-A1 Urea Channel Interactome
DescriptionThe urea channel Slc14a2 (or UT-A1) mediates vasopressin-regulated urea transport across the inner medullary collecting duct (IMCD). Previously, UT-A1 was found to present in a high molecular weight complex, suggesting UT-A1 is involved in certain protein-protein interactions. The present study sought to identify the proteins that interact with UT-A1 in this complex for a better understanding of how UT-A1 is regulated. Rat IMCD suspensions were treated with or without V2 receptor agonist, dDAVP, followed by in-cell crosslinking using BSOCOES and detergent solubilization. Immunoprecipitation using Dynabeads coated with UT-A1 specific antibody successfully pulled down the UT-A1 proteins. In-gel digestion protocol was carried out to prepare samples for liquid chromatographic mass spectrometry analysis of tryptic peptides using a Velos-Orbitrap mass spectrometer. The peptides passing stringent spectral quality thresholds were quantified (label-free) to identify those with (UTA-1 antibody/preimmune IgG) >4. A total of 128 UT-A1 interacting proteins were identified. Gene Ontology analysis maps the distribution of these proteins throughout major cell compartments: endoplasmic reticulum, Golgi, endosomes, cytosol and plasma membrane. Among them are four protein kinases (Cdc42bpb, Phkb, Camk2d, Mtor) that play roles in vasopressin-regulated phosphorylation of UT-A1. Non-label quantification was also performed to determine the stoichiometry of UT-A3 with UT-A1, the result does not support an oligomeric complex formation of UT-A1/A3. In conclusion, we have provided a refined list of UT-A1 binding proteins which can be useful for further analysis of the vasopressin signaling pathway in regulation of UT-A1 in IMCD.
HostingRepositoryPRIDE
AnnounceDate2019-11-08
AnnouncementXMLSubmission_2019-11-08_09:12:10.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterChung-Lin Chou
SpeciesList scientific name: Rattus norvegicus (Rat); NCBI TaxID: 10116;
ModificationListmonohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
InstrumentLTQ Orbitrap Elite
Dataset History
RevisionDatetimeStatusChangeLog Entry
02016-01-21 01:39:19ID requested
12019-11-08 09:12:12announced
Publication List
Chou CL, Hwang G, Hageman DJ, Han L, Agrawal P, Pisitkun T, Knepper MA, Identification of UT-A1- and AQP2-interacting proteins in rat inner medullary collecting duct. Am J Physiol Cell Physiol, 314(1):C99-C117(2018) [pubmed]
Keyword List
curator keyword: Biomedical
submitter keyword: asopressin, interacting protein, protein kinase, stoichiometry, mass spectrometry, kidney.
Contact List
Mark A. Knepper
contact affiliationEpithelial Systems Biology Laboratory, National Heart, Lung, and Blood Institute, National Institutes of Health
contact emailknep@helix.nih.gov
lab head
Chung-Lin Chou
contact affiliationNational Institutes of Health
contact emailchouj@nhlbi.nih.gov
dataset submitter
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