一种与亮抑酶肽样肽醛、甘氨酰甘氨酰-L-精氨醛偶联的琼脂糖衍生物及其作为胰蛋白酶亲和吸附剂的用途。

A sepharose derivative coupled with a leupeptin-like peptide aldehyde, glycylglycyl-L-argininal, and its use as an affinity adsorbent for trypsin.

作者信息

Nishikata M, Kasai K, Ishii S

出版信息

Biochim Biophys Acta. 1981 Aug 13;660(2):256-61. doi: 10.1016/0005-2744(81)90168-6.

Abstract

A Sepharose derivative containing a peptide aldehyde, glycylglycyl-L-argininal, the structure of which resembles that of leupeptin was prepared. It was a strong affinity adsorbent for trypsin (EC 3.4.21.4). Bovine trypsin showed higher affinity for this adsorbent at the optimum pH of catalysis (8.2) than at lower pH (5.0). This observation was in good agreement with the pH dependence of the interaction of leupeptin and trypsin (Kuramochi, H., Nakata, H. and Ishii, S. (1979) J. Biochem. 86, 1403-1410). Streptomyces griseus trypsin was also adsorbed while trypsinogen, alpha-chymotrypsin and TLCK-trypsin were not adsorbed. Though anhydrotrypsin, in which Ser-183 is converted to dehydroalanine, was not adsorbed, carbamoylmethylated (His-46) trypsin was adsorbed. Ser-183 proved to be essential for the binding. This adsorbent can also be used as a good tool to study the mechanism of action of leupeptin.

摘要

制备了一种含有肽醛甘氨酰甘氨酰-L-精氨醛的琼脂糖衍生物，其结构与亮抑酶肽相似。它是胰蛋白酶（EC 3.4.21.4）的强亲和吸附剂。在催化的最佳pH（8.2）下，牛胰蛋白酶对该吸附剂的亲和力高于在较低pH（5.0）时。这一观察结果与亮抑酶肽和胰蛋白酶相互作用的pH依赖性很好地吻合（仓持博、中田浩和石井秀夫（1979年）《生物化学杂志》86卷，1403 - 1410页）。灰色链霉菌胰蛋白酶也被吸附，而胰蛋白酶原、α-胰凝乳蛋白酶和甲苯磺酰-L-赖氨酸氯甲基酮处理的胰蛋白酶未被吸附。虽然其中Ser-183转化为脱氢丙氨酸的脱水胰蛋白酶未被吸附，但氨甲酰甲基化（His-46）的胰蛋白酶被吸附。结果证明Ser-183对结合至关重要。这种吸附剂也可作为研究亮抑酶肽作用机制的良好工具。