PXD002772

PXD002772 is an original dataset announced via ProteomeXchange.

Title	Structural determinants of heparin–transforming growth factor-β1 interactions and their effects on signalling
Description	Transforming growth factor-ß1 (TGF-ß1, Uniprot: P01137) is a heparin-binding protein that has been implicated in a number of physiological processes, including the initiation of chondrogenesis by human mesenchymal stem cells (hMSCs). Here, we identify the molecular features in the protein and in heparin required for binding and their effects on the potentiation of TGF-ß1’s activity on hMSCs. Using a proteomics “Protect and Label” approach, lysines K291, K304, K309, K315, K338, K373, K375 and K388 were identified as being directly involved in binding heparin. Competition assays in an optical biosensor demonstrated that TGF-ß1 does require N- and 6-O-sulfate groups for binding but that 2-O-sulfate groups are unlikely to underpin the interaction. Heparin-derived oligosaccharides as short as degree of polymerization (dp) 4 have a weak ability to compete for TGF-ß1 binding to heparin, which increases with the length of the oligosaccharide to reach a maximum between dp18 and dp24. In cell-based assays, heparin, 2-O-, 6-O- and N-desulfated re-N-acetylated heparin and oligosaccharides 14–24 saccharides (dp14–24) in length all increased the phosphorylation of SMAD2 after 6 h of stimulation with TGF-ß1. The results provide the structural basis for a model of heparin/heparan sulfate binding to TGF-ß1 and demonstrate that the features in the polysaccharide required for binding are not identical to those required for sustaining the signaling by TGF-ß1 in hMSCs.
HostingRepository	PRIDE
AnnounceDate	2015-09-01
AnnouncementXML	Submission_2015-09-01_01:36:43.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD002772
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Rachel Li
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	Oxidation; Acetyl; Carbamidomethyl; Biotin
Instrument	LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2015-08-21 00:42:30	ID requested
⏵ 1	2015-09-01 01:36:44	announced

Lee J, Wee S, Gunaratne J, Chua RJ, Smith RA, Ling L, Fernig DG, Swaminathan K, Nurcombe V, Cool SM, 1 interactions and their effects on signaling. Glycobiology, 25(12):1491-504(2015) [pubmed]

curator keyword: Biomedical

submitter keyword: heparan sulfate, heparin, proteoglycans, transforming growth factor-β1

Victor Nurcombe; Simon COOL
contact affiliation	Glycotherapeutics, IMB, A-star, Singapore
contact email	simon.cool@imb.a-star.edu.sg
lab head
Rachel Li
contact affiliation	IMCB
contact email	hxli@imcb.a-star.edu.sg
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/09/PXD002772

PRIDE project URI

• PRIDE
  1. PXD002772
     1. Label: PRIDE project
     2. Name: Structural determinants of heparin–transforming growth factor-β1 interactions and their effects on signalling