Kumazaki T, Kasai I, Ishii S
J Biochem. 1976 Apr;79(4):749-55. doi: 10.1093/oxfordjournals.jbchem.a131127.
An affinity adsorbent for trypsin [EC 3.4.21.4] (GGA Sepharose) was prepared. Glycylglycyl-L-arginine (GGA) was synthesized by a simple procedure and was immobilized on agarose gel. This adsorbent proved to have essentially the same characteristics as AP Sepharose, which is an affinity adsorbent containing tryptic peptides of protamine (1). GGS Sepharose was specific for native trypsin and had a stronger affinity at lower pH's (6-5) than at the optimum pH of trypsin action (8.2). It also proved to be suitable for analytical experiments because of its relatively weak affinity. By comparison of the elution profiles of trypsin from GGA Sepharose under various conditions, the nature of the interaction of trypsin with the adsorbent could be studied. It was found that alpha- and beta-trypsin could be distinguished. In the presence of arginine and N-substitute arginines, the elution of trypsin was accelerated. From the extents of the accelerating effects, the affinities of these compouunds could be compared.
制备了一种用于胰蛋白酶[EC 3.4.21.4]的亲和吸附剂（甘氨酰甘氨酰-L-精氨酸琼脂糖凝胶，GGA Sepharose）。通过简单的方法合成了甘氨酰甘氨酰-L-精氨酸（GGA），并将其固定在琼脂糖凝胶上。该吸附剂被证明具有与精蛋白胰蛋白酶肽亲和吸附剂（AP Sepharose）基本相同的特性(1)。甘氨酰甘氨酰-L-精氨酸琼脂糖凝胶（GGS Sepharose）对天然胰蛋白酶具有特异性，并且在较低pH值（6 - 5）下比在胰蛋白酶作用的最佳pH值（8.2）时具有更强的亲和力。由于其相对较弱的亲和力，它也被证明适用于分析实验。通过比较在各种条件下胰蛋白酶从甘氨酰甘氨酰-L-精氨酸琼脂糖凝胶上的洗脱曲线，可以研究胰蛋白酶与吸附剂相互作用的性质。发现可以区分α-胰蛋白酶和β-胰蛋白酶。在精氨酸和N-取代精氨酸存在下，胰蛋白酶的洗脱加速。从加速作用的程度，可以比较这些化合物的亲和力。
