Determination of the association constant of monovalent mode protein-sugar interaction by capillary zone electrophoresis.
Protein-sugar interaction was observed by capillary zone electrophoresis, using a few beta-galactose-specific lectins and lactobionic acid as protein and sugar models, respectively. The lectin peaks were retarded in a concentration-dependent manner by addition of lactobionic acid in a carrier, and association constants of monovalent mode interactions could be obtained from t1 (migration time of protein), t2 (migration time of complex, obtainable as the migration time at the plateau) and the slope of the (t-t1)-1 vs. [S]-1 plots, where t and [S] are the migration of protein in the presence of lactobionic acid and the concentration of lactobionic acid, respectively. The values for Ricinus communis agglutinin, peanut agglutinin and soy bean agglutinin at pH 6.8 were 3.3 . 10(3), 9.1 . 10(2) and 1.1 . 10(2)1 mol-1, respectively. This method required only small amounts of protein samples and was reproducible. The amount of the sugar could be minimized under the conditions that the carrier was a buffer containing the sugar whereas the electrode solutions consisted only of the buffer.
Honda
S
S
Faculty of Pharmaceutical Sciences, Kinki University, Higashi-osaka, Japan.
Taga
A
A
Suzuki
K
K
Suzuki
S
S
Kakehi
K
K
D002241:Carbohydrates / Q000737:chemistry; D003956:Dialysis; D004187:Disaccharides / Q000032:analysis; D004586:Electrophoresis; D037102:Lectins / Q000032:analysis; D011506:Proteins / Q000737:chemistry*
D002241:Carbohydrates; D004187:Disaccharides; D037102:Lectins; D011506:Proteins