Publication Details

Category Text Publication
Reference Category Journals
DOI 10.1515/hsz-2021-0167
Licence creative commons licence
Title (Primary) Identification of intracellular glycosaminoglycan-interacting proteins by affinity purification mass spectrometry
Author Großkopf, H.; Vogel, S.; Müller, C.D.; Köhling, S.; Dürig, J.-N.; Möller, S.; Schnabelrauch, M.; Rademann, J.; Hempel, U.; von Bergen, M.; Schubert, K.
Source Titel Biological Chemistry
Year 2021
Department MOLSYB
Volume 402
Issue 11
Page From 1427
Page To 1440
Language englisch
Topic T9 Healthy Planet
Keywords GAG-biotin; hBMSC; LC-MS/MS; pull-down approach; SaOS2/osteoblast cell line; sulfated glycosaminoglycan derivatives
Abstract Glycosaminoglycans (GAGs) are essential functional components of the extracellular matrix (ECM). Artificial GAGs like sulfated hyaluronan (sHA) exhibit pro-osteogenic properties and boost healing processes. Hence, they are of high interest for supporting bone regeneration and wound healing. Although sulfated GAGs (sGAGs) appear intracellularly, the knowledge about intracellular effects and putative interaction partners is scarce. Here we used an affinity-purification mass spectrometry-based (AP-MS) approach to identify novel and particularly intracellular sGAG-interacting proteins in human bone marrow stromal cells (hBMSC). Overall, 477 proteins were found interacting with at least one of four distinct sGAGs. Enrichment analysis for protein localization showed that mainly intracellular and cell-associated interacting proteins were identified. The interaction of sGAG with α2-macroglobulin receptor-associated protein (LRPAP1), exportin-1 (XPO1), and serine protease HTRA1 (HTRA1) was confirmed in reverse assays. Consecutive pathway and cluster analysis led to the identification of biological processes, namely processes involving binding and processing of nucleic acids, LRP1-dependent endocytosis, and exosome formation. Respecting the preferentially intracellular localization of sGAG in vesicle-like structures, also the interaction data indicate sGAG-specific modulation of vesicle-based transport processes. By identifying many sGAG-specific interacting proteins, our data provide a resource for upcoming studies aimed at molecular mechanisms and understanding of sGAG cellular effects.
Persistent UFZ Identifier
Großkopf, H., Vogel, S., Müller, C.D., Köhling, S., Dürig, J.-N., Möller, S., Schnabelrauch, M., Rademann, J., Hempel, U., von Bergen, M., Schubert, K. (2021):
Identification of intracellular glycosaminoglycan-interacting proteins by affinity purification mass spectrometry
Biol. Chem. 402 (11), 1427 - 1440 10.1515/hsz-2021-0167