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Obermann, J. ; Priglinger, C.S.* ; Merl-Pham, J. ; Geerlof, A. ; Priglinger, S.G.* ; Götz, M. ; Hauck, S.M.

Proteome-wide identification of glycosylation-dependent interactors of Galectin-1 and Galectin-3 on mesenchymal retinal pigment epithelial cells.

Mol. Cell. Proteomics 16, 1528-1546 (2017)
Publ. Version/Full Text Postprint Research data Research data DOI PMC
Open Access Green
Identification of interactors is a major goal in cell biology. Not only protein-protein but also protein-carbohydrate interactions are of high relevance for signal transduction in biological systems. Here we aim to identify novel interacting binding partners for the β-galactoside-binding proteins Galectin-1 (Gal-1) and Galectin-3 (Gal-3) relevant in the context of the eye disease proliferative vitreoretinopathy (PVR). PVR is one of the most common failures after retinal detachment surgeries and is characterized by the migration, adhesion and epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells (RPE) and the subsequent formation of sub- and epiretinal fibrocellular membranes. Gal-1 and Gal-3 bind in a dose- and carbohydrate-dependent manner to mesenchymal RPE cells and inhibit cellular processes like attachment and spreading. Yet knowledge about glycan-dependent interactors of Gal-1 and Gal-3 on RPE cells is very limited, although this is a prerequisite for unravelling the influence of galectins on distinct cellular processes in RPE cells. We identify here 131 Gal-3 and 15 Gal-1 interactors by galectin pull-down experiments combined with quantitative proteomics. They mainly play a role in multiple binding processes and are mostly membrane proteins. We focused on two novel identified interactors of Gal-1 and Gal-3 in the context of PVR: the low-density lipoprotein receptor LRP1 and the platelet-derived growth factor receptor beta PDGFRB. Addition of exogenous Gal-1 and Gal-3 induced crosslinking with LRP1/PDGFRB and Integrin-β1 (ITGB1) on the cell surface of human RPE cells and induced ERK/MAPK and Akt signaling. Treatment with Kifunensine, an inhibitor of complex-type-N-glycosylation, weakened the binding of Gal-1 and Gal-3 to these interactors and prevented lattice formation. In conclusion, the identified specific glycoprotein ligands shed light into the highly specific binding of galectins to dedifferentiated RPE cells and the resulting prevention of PVR-associated cellular events.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Glycoproteins* ; Mass Spectrometry ; N-glycosylation ; Protein-protein Interactions* ; Quantification ; Epithelial-to-mesenchymal Transition ; Galectins ; Lattice Formation ; Retinal Pigment Epithelial Cells; Receptor-related Protein; Experimental Proliferative Vitreoretinopathy; Carbohydrate-recognition Domain; Galactoside-binding Lectins; Amyloid Precursor Protein; Rich Akt Substrate; Lipoprotein Receptor; Cross-linking; T-cells; Alpha-2-macroglobulin Receptor
ISSN (print) / ISBN 1535-9476
e-ISSN 1535-9484
Journal Molecular and Cellular Proteomics
Quellenangaben Volume: 16, Issue: 8, Pages: 1528-1546 Article Number: , Supplement: ,
Publisher American Society for Biochemistry and Molecular Biology
Publishing Place Bethesda
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) CF Metabolomics & Proteomics (CF-MPC)
Institute of Structural Biology (STB)
Institute of Stem Cell Research (ISF)