caa a22 4500 397555679 CHVBK 20180308164801.0 cr unu---uuuuu 161202e199612 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021534 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021534 Determination of the Affinity Constants of Pea Lectin for Neutral Sugars by Capillary Affinophoresis with a Monoligand Affinophore [Elektronische Daten] [Kiyohito Shimura, Ken-ichi Kasai] Affinophoresis is a type of affinity electrophoresis in which an affinophore, a conjugate of an affinity ligand and a multiply charged soluble matrix, causes a change in migration velocity of proteins which have a specific affinity for the ligand. A monoligand affinophore bearing a mannoside was prepared by coupling iodoacetylated p-aminophenyl α-D-mannoside to the free thiol group of N-succinylated glutathione, and used for the affinophoresis of pea lectin in a capillary. The electrophoretic mobility of pea lectin towards the anode increased in the presence of the affinophore as a function of its concentration in a manner that is described by the equation for affinity electrophoresis. Analysis of the suppression of the affinophoresis on the addition of neutral sugars to the system allowed the determination of the dissociation constants of the lectin for these neutral sugars. The dissociation constants obtained on affinophoresis agreed well with the values in the literature. The preparation of a monoligand affinophore for ligands bearing an amino group should facilitate the application of this type of microscale analysis (0.14 ng of protein for each run) to protein ligand interactions. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence affinity electrophoresis nationallicence capillary electrophoresis nationallicence lectin nationallicence ligand binding nationallicence molecular recognition nationallicence Shimura Kiyohito Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko, Kanagawa 199-01 aut Kasai Ken-ichi Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko, Kanagawa 199-01 aut The Journal of Biochemistry Oxford University Press 120/6(1996-12), 1146-1152 0021-924X 120:6<1146 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021534 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021534 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Shimura Kiyohito Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko, Kanagawa 199-01 aut NATIONALLICENCE 700 1- Kasai Ken-ichi Department of Biological Chemistry, Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko, Kanagawa 199-01 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/6(1996-12), 1146-1152 0021-924X 120:6<1146 1996 120 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford