PXD035472-1

PXD035472 is an original dataset announced via ProteomeXchange.

Title	PRM targeted mass spectrometry reveals the phosphorylation regulation of human bile acid transporter ASBT
Description	The 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.
HostingRepository	PanoramaPublic
AnnounceDate	2024-09-05
AnnouncementXML	Submission_2024-09-05_08:34:20.202.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	supported by repository but incomplete data and/or metadata
PrimarySubmitter	Thi Thao Nguyen
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	Phospho; Label:13C(6)15N(2)
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2022-07-20 21:54:24	ID requested
⏵ 1	2024-09-05 08:34:20	announced

no publication

submitter keyword: ASBT, PRM, Phosphorylation stochiometry

Maureen Kane
contact affiliation	University of Maryland, School of Pharmacy
contact email	mkane@rx.umaryland.edu
lab head
Thi Thao Nguyen
contact affiliation	University of Maryland, School of Pharmacy
contact email	thaolibra2010@gmail.com
dataset submitter

Panorama Public dataset URI