Kohlmiller N A, Howard J B
J Biol Chem. 1979 Aug 10;254(15):7302-8.
The carboxymethylated alpha subunit of protocatechuate 3,4-dioxygenase was digested with trypsin. The 14 tryptic peptides were isolated by ion exchange chromatography on DEAE-Sephadex and by gel filtration chromatography. Automated Edman degradation and carboxypeptidase Y and B digestion were used to establish the sequence of these peptides. Further fragmentation of two tryptic peptides, T3 and T5, by Staphylococcus aureus protease and cyanogen bromide, respectively, was necessary to complete the sequences. The tryptic peptides accounted for a minimum of 199 residues out of a total of 202 residues predicted by amino acid analysis.
将原儿茶酸3,4-双加氧酶的羧甲基化α亚基用胰蛋白酶消化。通过在DEAE-葡聚糖上进行离子交换色谱和凝胶过滤色谱分离出14个胰蛋白酶肽段。使用自动Edman降解以及羧肽酶Y和B消化来确定这些肽段的序列。为了完成序列，分别用金黄色葡萄球菌蛋白酶和溴化氰对两个胰蛋白酶肽段T3和T5进行进一步裂解是必要的。氨基酸分析预测该蛋白质共有202个残基，这些胰蛋白酶肽段至少占其中的199个残基。
