PXD050256

PXD050256 is an original dataset announced via ProteomeXchange.

Title	Analysis of endogenous NOTCH1 from POFUT1 S162L patient fibroblasts reveals the importance of the O-fucose modification on EGF12 in human development
Description	NOTCH1 (N1) is a transmembrane receptor interacting with membrane-tethered ligands on opposing cells that mediate the direct cell-cell interaction necessary for many cell fate decisions. Protein O-fucosyltransferase 1 (POFUT1) adds O-fucose to Epidermal Growth Factor (EGF)-like repeats in the NOTCH extracellular domain, which is required for signaling activation and trafficking. We previously showed that POFUT1 S162L caused a 90% loss of POFUT1 activity and global developmental defects in a patient; however, the mechanism by which POFUT1 contributes to these symptoms is still unclear. Compared to controls, POFUT1 S162L patient fibroblast cells had an equivalent amount of N1 on the cell surface but showed a 60% reduction of DLL1 ligand binding and a 70% reduction in JAG1 ligand binding. To determine if the reduction of O-fucose on N1 in POFUT1 S162L patient fibroblasts is the cause of these effects, we immunopurified endogenous N1 from control and patient fibroblasts and analyzed O-fucosylation using mass spectral glycoproteomics methods. N1 EGF8 to EGF12 comprise the ligand binding domain, and O-fucose on EGF8 and EGF12 physically interact with ligands to enhance affinity. Glycoproteomics of N1 from POFUT1 S162L patient fibroblasts showed WT fucosylation levels at all sites analyzed except for a large decrease at EGF9 and the complete absence of O-fucose at EGF12. Since the loss of O-fucose on EGF12 has significant effects on N1 activity, this may explain the symptoms observed in the POFUT1 S162L patient.
HostingRepository	PRIDE
AnnounceDate	2024-08-02
AnnouncementXML	Submission_2024-08-02_05:15:40.220.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Kenjiroo Matsumoto
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	carboxylated residue; monohydroxylated residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2024-02-29 19:34:51	ID requested
⏵ 1	2024-08-02 05:15:40	announced

10.1101/2024.04.09.588484;

Matsumoto K, Luther KB, Haltiwanger RS, -fucose modification on EGF12 in human development. bioRxiv, ():(2024) [pubmed]

submitter keyword: NOTCH1, mass spectrometry, O-fucose, POFUT1

Robert Haltiwanger
contact affiliation	Complex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia, USA.
contact email	rhalti@uga.edu
lab head
Kenjiroo Matsumoto
contact affiliation	Complex Carbohydrate Research Center
contact email	kenjiroo.matsumoto@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD050256
     1. Label: PRIDE project
     2. Name: Analysis of endogenous NOTCH1 from POFUT1 S162L patient fibroblasts reveals the importance of the O-fucose modification on EGF12 in human development