A heparin-binding peptide from human serum amyloid P component characterized by affinity capillary electrophoresis

Electrophoresis
N H Heegaard

Abstract

Affinity capillary electrophoresis (CE) was used for a detailed characterization of the binding between heparin and a peptide isolated from the heparin-binding serum protein amyloid P component (SAP). The peptide corresponds to a tryptic fragment (T3) comprising amino acids 14-38 of SAP. By including ligands in the electrophoresis buffer various glycosaminoglycans could be screened for binding of T3 using one sample aliquot. The binding was found to be highly specific for heparin and heparin fragments down to tetramers and appeared strongest at a slightly alkaline pH while no binding could be demonstrated with heparan sulfate, chondroitin sulfate, desulfated heparin, mannose 6-phosphate and phosphotyrosine. The T3-heparin complexes were sufficiently stable to perform quantitative measurements of the binding using preequilibration of samples prior to a CE-mediated separation of bound and free T3-peptide. Plots based on quantitation of analyte peaks corresponding to free and complexed T3 yielded a dissociation constant of 1.5 microM for the interaction with heparin. The results indicate that a specific subfraction of the heparin molecules is active in binding interactions with the peptide. The affinity CE approach proved to be us...Continue Reading

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Citations

Dec 20, 2002·Angewandte Chemie·Ishan Capila, Robert J Linhardt
May 31, 2001·Electrophoresis·K L Rundlett, D W Armstrong
Feb 22, 2002·Biomedical Chromatography : BMC·Wenjun MaoRobert J Linhardt
Nov 25, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·C BertucciV Andrisano
Apr 10, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Yoshihide Tanaka, Shigeru Terabe
Jun 1, 2005·Journal of Pharmaceutical and Biomedical Analysis·Aiye LiangBingcheng Lin
Jun 12, 2009·Electrophoresis·Niels H H Heegaard
May 30, 2006·Electrophoresis·Jesper Ostergaard, Niels H H Heegaard
Mar 29, 2002·Electrophoresis·Niels H H HeegaardDavid D Y Chen
Dec 22, 1999·Electrophoresis·V Kasicka

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