Determination of dissociation constants for a heparin-binding domain of amyloid precursor protein and heparins or heparan sulfate by affinity capillary electrophoresis.
Dynamic affinity capillary electrophoresis (ACE) was used for determining the binding constants between heparin-like glycosaminoglycans and the (96-110) heparin-binding domain of amyloid precursor protein (APP). The migration time shift of the (96-110) APP peptide was monitored as the concentration of heparin was increased in the background electrolyte. The compounds investigated included low-molecular-weight heparin, porcine mucosa heparin, and heparan sulfate. Change in mobility as a function of glycosaminoglycan concentration was plotted using both linear regression (Scatchard analysis) and nonlinear regression. Dissociation constants (K(d)) were determined and compared for both sets of analyses with the low-molecular-weight heparin giving the most reproducible results and best fit with a K(d) value of 3.9 microM.
McKeon
Jocelyn
J
Chemistry Department, West Virginia University, Morgantown, WV, USA.
Holland
Lisa A
LA
D016564:Amyloid beta-Protein Precursor / Q000737:chemistry*; D019075:Electrophoresis, Capillary; D006025:Glycosaminoglycans / Q000737:chemistry*; D006493:Heparin / Q000737:chemistry*; D006497:Heparan Sulfate / Q000737:chemistry*; D006801:Humans; D011485:Protein Binding; D017434:Protein Structure, Tertiary
D016564:Amyloid beta-Protein Precursor; D006025:Glycosaminoglycans; D006493:Heparin; D006497:Heparan Sulfate