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 and hence are of high interest for supporting wound healing. Although sulfated GAGs (sGAGs) are internalized into cells, knowledge of intracellular effects and 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 from human bone marrow stromal cells (hBMSC). Overall, 478 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. Only a limited fraction of about 10 % was annotated for extracellular regions. The interactions of the 2-macroglobulin receptor-associated protein (LRPAP1), exportin-1 (XPO1), and Serine protease HTRA1 (HTRA1) were confirmed in reverse assays.