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Determination of dissociation constants for a heparin‐binding domain of amyloid precursor protein and heparins or heparan sulfate by affinity capillary electrophoresis
Author(s) -
McKeon Jocelyn,
Holland Lisa A.
Publication year - 2004
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200405878
Subject(s) - heparan sulfate , chemistry , heparin , capillary electrophoresis , dissociation constant , glycosaminoglycan , scatchard plot , affinity electrophoresis , electrophoresis , chromatography , peptide , dissociation (chemistry) , low molecular weight heparin , biochemistry , binding site , affinity chromatography , enzyme , receptor , organic chemistry
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 μ M .