Belozersky M A, Dunaevsky Y E, Voskoboynikova N E
A.N. Belozersky Laboratory of Molecular Biology and Bioorganic Chemistry, Moscow State University, U.S.S.R.
Biochem J. 1990 Dec 15;272(3):677-82. doi: 10.1042/bj2720677.
A homogeneous preparation of metalloproteinase, purified 1000-fold, was obtained from buckwheat (Fagopyrum esculentum) seeds. The Mr of the enzyme, determined by SDS/PAGE, was 34,000 (it was 39,000 by gel chromatography). Its pH optimum was 8.0-8.2 with 13 S globulin, from buckwheat seeds, as substrate. Atomic-absorption spectroscopy revealed the presence of one Zn2+ ion per enzyme molecule. The enzyme was completely inhibited by EDTA (1 mM), zincone (1 mM) and 1, 10-phenanthroline (1 mM). The metalloproteinase performed limited proteolysis of the following seed storage proteins: 13 S globulin from buckwheat seeds and 11 S globulin from soybean (Glycine max) seeds. It hydrolysed three peptide bonds formed by the amino groups of Leu15, Tyr16 and Phe25 in the oxidized B-chain of insulin. In its main properties the enzyme is similar to metalloproteinases of animal and bacterial origin.
从荞麦（苦荞麦）种子中获得了一种纯化了1000倍的金属蛋白酶均质制剂。通过SDS/PAGE测定，该酶的相对分子质量为34,000（凝胶色谱法测定为39,000）。以荞麦种子中的13S球蛋白为底物时，其最适pH为8.0 - 8.2。原子吸收光谱显示每个酶分子含有一个Zn2+离子。该酶被EDTA（1 mM）、锌酮（1 mM）和1,10 - 菲咯啉（1 mM）完全抑制。这种金属蛋白酶对以下种子贮藏蛋白进行有限的蛋白水解：荞麦种子中的13S球蛋白和大豆（大豆）种子中的11S球蛋白。它水解了胰岛素氧化B链中由Leu15、Tyr16和Phe25的氨基形成的三个肽键。该酶的主要性质与动物和细菌来源的金属蛋白酶相似。
