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PXD035472-1

PXD035472 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitlePRM targeted mass spectrometry reveals the phosphorylation regulation of human bile acid transporter ASBT
DescriptionThe human apical bile acid transporter (hASBT) in the distal ileum reabsorbs bile acids and is responsible for transporting approximately 95 % of bile acids back to liver for recycling in the enterohepatic pathway. ASBT activity has been linked to multiple disease states, including Crohn’s disease, hypercholesterolemia, cholestasis, and type 2 diabetes. ASBT activity is regulated at the post-translational level by glycosylation, ubiquitination, and S-acylation, which control its translocation to the cell surface and protein stability. Although biochemical studies have suggested that phosphorylation of serine, threonine, and tyrosine may play a role in the regulation of ASBT function, no study has determined where phosphorylation of ASBT occurs and how its phosphorylation level correlates with its activity. In this study, we developed a workflow using parallel reaction monitoring (PRM) targeted mass spectrometry to determine and quantify the stoichiometry of ASBT phosphorylation in the presence of various kinase inhibitors and activators. Our findings provide the first evidence of ASBT phosphorylation at multiple sites (Thr330, Ser334, and Ser 335), with Ser 335 being the most abundant phosphosite.Futhermore, we demonstrate that the phosphorylation level of Serine 335 coincides with the bile acid uptake activity of ASBT. Finally, we discovered that PKCalpha, but not other PKC isoforms, regulates ASBT phosphorylation at Ser 335. Taken together, our findings establish the molecular basis of phosphorylation-mediated regulation of ASBT which may include novel therapeutic targets for managing ASBT-linked disease.
HostingRepositoryPanoramaPublic
AnnounceDate2024-09-05
AnnouncementXMLSubmission_2024-09-05_08:34:20.202.xml
DigitalObjectIdentifier
ReviewLevelNon peer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportsupported by repository but incomplete data and/or metadata
PrimarySubmitterThi Thao Nguyen
SpeciesList scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationListPhospho; Label:13C(6)15N(2)
InstrumentQ Exactive HF
Dataset History
RevisionDatetimeStatusChangeLog Entry
02022-07-20 21:54:24ID requested
12024-09-05 08:34:20announced
Publication List
no publication
Keyword List
submitter keyword: ASBT, PRM, Phosphorylation stochiometry
Contact List
Maureen Kane
contact affiliationUniversity of Maryland, School of Pharmacy
contact emailmkane@rx.umaryland.edu
lab head
Thi Thao Nguyen
contact affiliationUniversity of Maryland, School of Pharmacy
contact emailthaolibra2010@gmail.com
dataset submitter
Full Dataset Link List
Panorama Public dataset URI