Yamada M, Tashiro M, Yamaguchi H, Yamada H, Ibuki F
J Biochem. 1976 Dec;80(6):1293-7. doi: 10.1093/oxfordjournals.jbchem.a131401.
The reactive site peptide bond of the eggplant inhibitor against trypsin [EC 3.4.21.4] was identified by chemical modifications with 1,2-cyclohexanedione, 2,4,6-trinitrobenzenesulfonic acid, acetic anhydride and glyoxal, and by sequential treatments with trypsin and carboxypeptidase B [EC 3.4.12.3]. The inhibitor was significantly inactivated by chemical modifications of arginine residues, but was not affected by lysine modifications. Free arginine was released from the trypsin-modified inhibitor by carboxypeptidase B digestion, accompanied by a marked loss of inhibitory activity. A serine residue was newly exposed at the N-terminal amino acid of the inhibitor after modification with trypsin. The reactive site of the inhibitor against trypsin was concluded to be an arginylseryl bond. The inhibitor was completely inactivated by full reduction of its disulfide bonds.
通过用1,2 - 环己二酮、2,4,6 - 三硝基苯磺酸、乙酸酐和乙二醛进行化学修饰，以及用胰蛋白酶和羧肽酶B [EC 3.4.12.3] 进行顺序处理，确定了茄子胰蛋白酶抑制剂 [EC 3.4.21.4] 的反应位点肽键。精氨酸残基的化学修饰使该抑制剂显著失活，但赖氨酸修饰对其没有影响。羧肽酶B消化使胰蛋白酶修饰后的抑制剂释放出游离精氨酸，同时抑制活性显著丧失。用胰蛋白酶修饰后，抑制剂的N端氨基酸处新暴露了一个丝氨酸残基。得出该抑制剂对胰蛋白酶的反应位点是一个精氨酰丝氨酸键。该抑制剂的二硫键完全还原后会完全失活。
