Sulfated Hyaluronan Alters the Interaction Profile of TIMP-3 with the Endocytic Receptor LRP-1 Clusters II and IV and Increases the Extracellular TIMP-3 Level of Human Bone Marrow Stromal Cells

Abstract

Sulfated glycosaminoglycans (sGAGs) modulate cellular processes via their interaction with extracellular matrix (ECM) proteins. We revealed a direct binding of tissue inhibitor of metalloproteinase-3 (TIMP-3) to the endocytic receptor low-density lipoprotein receptor-related protein (LRP-1) clusters II and IV using surface plasmon resonance. Sulfated hyaluronan (sHA) and chondroitin sulfate (sCS) derivatives interfered with TIMP-3/LRP-1 complex formation in a sulfation-dependent manner stronger than heparin. Electrostatic potential calculations suggested a competition between negatively charged GAGs and highly negatively charged complement-like domains of LRP-1 for the binding to a positively charged area of TIMP-3 as an underlying mechanism. In vitro studies revealed increased amounts of pericellular TIMP-3 in the presence of sHA as a consequence of the blocked protein uptake. GAG derivatives as part of biomaterials might post-translationally modulate TIMP-3 levels stronger than native GAGs, thus exhibiting catabolic effects on the ECM, which could prevent extensive pathological matrix degradation and promote wound healing.

Sulfation Patterns of Saccharides and Heavy Metal Ion Binding

“…Am ore profounda nalysis will be required to elucidate the exact contribution of sulfation to electrostatic and conformational changes of oligosaccharides that leads to the observedh eavy metal ion preferences.O ur sensors provide us with valuable hints regardingt he effect of sulfation on metal binding preferencest hat adds to the current knowledge on the interaction of sulfated saccharides with their environment. [32,34] Our resultss uggestt hat the effect of sulfation on the metal binding cannot be simplified merely to the number of sulfate groups. The results indicatet hat the system is not as imple ion-exchanger but that it is more com-plex and that the distinctive sulfationp attern of saccharides is crucial for modulating the metal-binding affinities, perhaps throught he formation of asecondary coordination sphere.…”

“…Furthermore, we clearly observed that al arge number of sulfate groups interfere with the chelation ability.T hisp roves that simply "loading" ag lycan with sulfatesd oes not necessarily provide preference to metal ions and that the pattern is indeed crucial. [32,34] Our resultss uggestt hat the effect of sulfation on the metal binding cannot be simplified merely to the number of sulfate groups. The effect of sulfation on the conformation of furanosides and GAG was previously reported.…”

“…[2,3] Alternatively, artificial sulfated polymers such as polystyrene sulfonate have been used as replacements for saccharides but these compounds lack the rich structural properties of the native entities. [32][33][34][35] sHA have been successfully used to study the interactions with GAG-binding sites of many regulatory proteins and for the design of artificial extracellular matrices with tailored functions. [27] Sulfated hyaluronic acids (sHA) are synthetic oligosaccharides derived from hyaluronica cid (HA) that largely resemble natives ulfated GAG such as chondroitin,d ermatan or heparan sulfate ( Figure 1A and C).…”

“…[28][29][30][31] They were used to study protein glycani nteractions, for biomedical applications,a nd they are well accepted as biologically relevant GAG relatedm odels. [32][33][34][35] sHA have been successfully used to study the interactions with GAG-binding sites of many regulatory proteins and for the design of artificial extracellular matrices with tailored functions. [36][37][38][39] Recently,w eh ave developed strategies to produce av ariety of defined sHA from readily available HA in larger quantities by using ac ombination of enzymatic and chemical methods.…”

Glycosaminoglycan and Proteoglycan Biotherapeutics in Articular Cartilage Protection and Repair Strategies: Novel Approaches to Visco‐supplementation in Orthobiologics

“…[222] GAGs also modulate the activity and activation of the lysosomal exopeptidase tripeptidyl-peptidase-I thus may affect the processing of peptide hormones and growth factors in-situ and tissue repair. [55,223] TIMP-3 is the only TIMP which has interactive properties with ECM components which localize it in tissues. [22] Interactions between GAGs and TIMP-3 suggest that this may improve TIMP-3 inhibitory properties against MMPs and be protective in repair environments.…”