Gladysheva I P, Sharafutdinov T Z, Larionova N I
Bioorg Khim. 1994 Mar;20(3):281-9.
Multiple forms of Bowman-Birk soybean inhibitor have for the first time been isolated from commercial soya flour and purified to homogeneity. Amino acid compositions and isoelectric points of the inhibitors were determined. The isolated inhibitors are shown to be related to classic (M 8000 Da, 2-II) and high molecular mass glycine-rich (M 17 000 Da, 3-II, 5-II) Bowman-Birk inhibitors. The inhibitor (2-II) was found to have two reactive sites and bind trypsin at one centre and alpha-chymotrypsin, cathepsin G and human leukocyte elastase at the other. Rate constants of the complex formation (ka) and complex dissociation (kd) were determined by following the kinetics of approaching to the steady state in a system including the enzyme, the substrate and various concentrations of the inhibitor.
首次从市售大豆粉中分离出多种形式的鲍曼-伯克大豆抑制剂，并将其纯化至同质。测定了抑制剂的氨基酸组成和等电点。结果表明，分离出的抑制剂与经典的（分子量8000道尔顿，2-II）和高分子量富含甘氨酸的（分子量17000道尔顿，3-II，5-II）鲍曼-伯克抑制剂有关。发现抑制剂（2-II）有两个反应位点，在一个中心结合胰蛋白酶，在另一个中心结合α-糜蛋白酶、组织蛋白酶G和人白细胞弹性蛋白酶。通过跟踪包括酶、底物和不同浓度抑制剂的系统中接近稳态的动力学，测定了复合物形成（ka）和复合物解离（kd）的速率常数。
