Interaction of soluble and surface‐bound heparin binding growth‐associated molecule with heparin

FEBS Letters

Fath, Melissa²  Kilpeläinen, Ilkka¹  Linhardt, Robert J.²  Rauvala, Heikki¹  Kinnunen, Tarja¹  VanderNoot, Victoria² 

[1] Laboratory of Molecular Neurobiology, Institute of Biotechnology and the Department of Neurosciences, University of Helsinki, Helsinki 00014, Finland;Division of Medicinal and Natural Products Chemistry and Department of Chemical and Biochemical Engineering, University of Iowa, Iowa City, IA 52242, USA

关键词: Heparin;    Heparin binding growth-associated molecule;    HB-GAM;    Pleiotrophin;    Interaction;    Isothermal titration calorimetry;    Surface plasmon resonance;   

DOI  :  10.1016/S0014-5793(99)00785-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The interaction of heparin with heparin binding growth-associated molecule (HB-GAM) was studied using isothermal titration calorimetry (ITC) and surface plasmon resonance (SPR). ITC studies showed that, in solution, heparin bound HB-GAM with a ΔH of −30 kcal/mole corresponding to a dissociation constant (K _d) of 460 nM. The stoichiometry of interaction was 3 moles of HB-GAM per mole of heparin, corresponding to a minimum heparin binding site for HB-GAM of 12–16 saccharide residues. Kinetic measurements of heparin interaction with HB-GAM made by SPR afforded a K _d of 4 nM, suggesting considerably tighter binding when HB-GAM was immobilized on a surface. Affinity chromatography of a sized mixture of heparin oligosaccharides, having a degree of polymerization (dp) of >14 saccharide units, on HB-GAM-Sepharose demonstrated that oligosaccharides having more than 18 saccharide residues showed the tightest interaction.

【 授权许可】

Unknown