Determination of the association constant of monovalent mode protein-sugar interaction by capillary zone electrophoresis.
Determination of the association constant of monovalent mode protein-sugar interaction by capillary zone electrophoresis.
复制标题
通过毛细管区带电泳测定单价模式蛋白-糖相互作用的缔合常数。
DOI：
10.1016/0021-9673(92)80134-g
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
K. Kakehi
中科院分区：
文献类型：
--
作者：
S. Honda;A. Taga;K. Suzuki;S. Suzuki;K. Kakehi
以β-半乳糖凝集素和乳糖酸分别作为蛋白质和糖的模型，用毛细管区带电泳法观察蛋白质-糖相互作用。通过在载体中加入乳糖酸，凝集素峰以浓度依赖性方式延迟，单价模式相互作用的缔合常数可以从t1（蛋白质的迁移时间），t2（复合物的迁移时间，可作为平台的迁移时间）和（t-t1）-1vs的斜率获得。[S]-1图，其中和[S]分别是存在乳糖酸和乳糖酸浓度时蛋白质的迁移。蓖麻凝集素、花生凝集素和大豆凝集素在pH 6.8时的测定值分别为3.3 · 103、9.1 ·102和1.1 · 1021 mol−1。该方法只需要少量的蛋白质样品，并且具有重现性。在载体是含有糖的缓冲液而电极溶液仅由缓冲液组成的条件下，糖的量可以最小化
Protein—sugar interaction was observed by capillary zone electrophoresis, using a few β-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 fromt1(migration time of protein),t2(migration time of complex, obtainable as the migration time at the plateau) and the slope of the (t-t1)−1vs. [S]−1plots, wheretand [S] are the migration of protein in the presence of lactobionic acid and the concentration of lactobionic acid, respectively. The values forRicinus communisagglutinin, peanut agglutinin and soy bean agglutinin at pH 6.8 were 3.3 · 103, 9.1 · 102and 1.1 · 1021 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