DETERMINATION OF THE A AFFINITY CONSTANTS OF PEA LECTIN FOR NEUTRAL SUGARS BY CAPILLARY AFFINOPHORESIS WITH A MONOLIGAND AFFINOPHORE

Authors
Citation
K. Shimura et K. Kasai, DETERMINATION OF THE A AFFINITY CONSTANTS OF PEA LECTIN FOR NEUTRAL SUGARS BY CAPILLARY AFFINOPHORESIS WITH A MONOLIGAND AFFINOPHORE, Journal of Biochemistry, 120(6), 1996, pp. 1146-1152
Citations number
22
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biology
Journal title
ISSN journal
0021-924X
Volume
120
Issue
6
Year of publication
1996
Pages
1146 - 1152
Database
ISI
SICI code
0021-924X(1996)120:6<1146:DOTAAC>2.0.ZU;2-P
Abstract
Affinophoresis is a type of affinity electrophoresis in which an affin ophore, a conjugate of an affinity ligand and a multiply charged solub le matrix, causes a change in migration velocity of proteins which hav e a specific affinity for the ligand. A monoligand affinophore bearing a mannoside was prepared by coupling iodoacetylated p-aminophenyl alp ha-D-mannoside to the free thiol group of N-succinylated glutathione, and used for the affinophoresis of pea lectin in a capillary, The elec trophoretic mobility of pea lectin towards the anode increased in the presence of the affinophore as a function of its concentration in a ma nner that is described by the equation for affinity electrophoresis. A nalysis of the suppression of the affinophoresis on the addition of ne utral sugars to the system allowed the determination of the dissociati on constants of the lectin for these neutral sugars, The dissociation constants obtained on affinophoresis agreed well with the values in th e 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 li gand interactions.