The Binding of Chondroitin Sulfate to Pleiotrophin/Heparin-binding Growth-associated Molecule Is Regulated by Chain Length and Oversulfated Structures
| Autor: | Nobuaki Maeda, Toshihiro Hata, Nobuna Fukazawa |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Time Factors
medicine.medical_treatment Oligosaccharides Chondroitin ABC Lyase Pleiotrophin Biochemistry High-performance liquid chromatography chemistry.chemical_compound medicine Animals Humans Chondroitin sulfate Shark cartilage Receptor Molecular Biology Chromatography High Pressure Liquid Chromatography Binding Sites Dose-Response Relationship Drug biology Heparin Chemistry Growth factor Chondroitin Sulfates Cell Biology Surface Plasmon Resonance Chromatography Ion Exchange Kinetics Cartilage Cross-Linking Reagents Proteoglycan Sharks biology.protein Cytokines Carrier Proteins Protein Binding Signal Transduction medicine.drug |
| Zdroj: | Journal of Biological Chemistry. 281:4894-4902 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m507750200 |
| Popis: | Pleiotrophin is an 18-kDa heparin-binding growth factor, which uses chondroitin sulfate (CS) proteoglycan, PTPzeta as a receptor. It has been suggested that the D-type structure (GlcA(2S)beta1-3GalNAc(6S)) in CS contributes to the high affinity binding between PTPzeta and pleiotrophin. Here, we analyzed the interaction of shark cartilage CS-D with pleiotrophin using a surface plasmon resonance biosensor to reveal the importance of D-type structure. CS-D was partially digested with chondroitinase ABC, and fractionated using a Superdex 75pg column. Theor =18-mer CS fractions showed significant binding to pleiotrophin, and the longer fractions had stronger affinity for pleiotrophin than the shorter ones. The approximately 46-mer CS fraction bound to densely immobilized pleiotrophin with high affinity (K(D) = approximately 30 nM), and the binding reactions fitted the bivalent analyte model. However, when the density of the immobilized pleiotrophin was lowered, the strength of affinity remarkably decreased (K(D) = approximately 2.5 microM), and the reactions no longer fitted the model and were considered to be monovalent binding. The 20 approximately 24-mer fractions showed low affinity binding to densely immobilized pleiotrophin (K(D) = 3 approximately 20 microM), which seemed to be monovalent. When approximately 22-mer CS oligosaccharides were fractionated by strong anion exchange HPLC, each fraction differed in affinity for pleiotrophin (K(D) = 0.36 approximately10 microM), and the affinity correlated with the amounts of D- and E- (GlcAbeta1-3GalNAc(4S,6S)) type oversulfated structures. These results suggest that the binding of pleiotrophin to CS is regulated by multivalency with CS approximately 20 mer as a unit and by the amounts of oversulfated structures. |
| Databáze: | OpenAIRE |
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