PXD029295

PXD029295 is an original dataset announced via ProteomeXchange.

Title	hepatocyte grwoth factor receptor (MET) LC-MSMS
Description	MET, the receptor for hepatocyte growth factor (HGF), is involved in wide variety of biological events and its aberrant activation is implicated in the pathogenesis of cancer. Especially, MET signaling is associated with resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs), key drugs in therapy of non-small cell lung cancer. MET has 11 potential N-glycosylation sites, however, the site-specific roles of N-glycans have not been elucidated. Here, we report that N-glycans regulate proteolytic processing of MET and HGF induced MET signaling, site specifically. N-glycosylation inhibitors suppressed processing and trafficking of endogenous MET in H1975 and EBC-1 lung cancer cells and exogenous MET in CHO cells. We purified the recombinant extracellular domain of human MET and determined the site-specific N-glycan structures and occupancy using nanoflow liquid chromatography-electrospray ionization mass spectrometry. The results indicated that most sites were fully glycosylated and the dominant population were complex-type which were rich in sialic acids and core fucoses. To examine the effects of N-glycan deletion of MET, we prepared endogenous-MET-knockout Flp-In CHO cells and transfected with series of N-glycan deletion mutants of MET. It was found that several N-glycans are implicated in processing of MET. It was also suggested that the N-glycans of -subunit of MET positively regulate HGF signaling, and the N-glycans of β-subunit negatively regulate HGF signaling. In all-N-glycan deletion mutant, processing and signaling were significantly suppressed. It was observed that the cell proliferation rate was suppressed in all-N-glycan deletion mutant, although the cell surface expression levels of MET were not completely diminished. We examined the HGF biding affinity for the recombinant extracellular domain of MET and found that peptide N-glycosidase F treatment did not affect the ligand binding affinity. It was suggested that N-glycans of MET affect the status and the function of the receptor site-specifically.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:44:44.515.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Naoki Fujitani
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2021-10-22 06:57:41	ID requested
1	2023-04-07 03:47:10	announced
⏵ 2	2023-11-14 08:44:47	announced	2023-11-14: Updated project metadata.

Saitou A, Hasegawa Y, Fujitani N, Ariki S, Uehara Y, Hashimoto U, Saito A, Kuronuma K, Matsumoto K, Chiba H, Takahashi M, N-glycosylation regulates MET processing and signaling. Cancer Sci, 113(4):1292-1304(2022) [pubmed]

submitter keyword: glycosylation,hepatocyte grwoth factor receptor (MET)

Motoko Takahashi
contact affiliation	Department of Biochemistry, Sapporo Medical University School of Medicine
contact email	takam@sapmed.ac.jp
lab head
Naoki Fujitani
contact affiliation	Sapporo Medical University
contact email	fujitani@sapmed.ac.jp
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD029295
     1. Label: PRIDE project
     2. Name: hepatocyte grwoth factor receptor (MET) LC-MSMS