固定在琼脂糖凝胶上的杏仁糖肽酶消化去糖基化的高峰淀粉酶A载脂蛋白的分离与鉴定。

Isolation and characterization of Taka-amylase A apoprotein deglycosylated by digestion with almond glycopeptidase immobilized on Sepharose.

作者信息

Takahashi N, Toda H, Nishibe H, Yamamoto K

出版信息

Biochim Biophys Acta. 1982 Oct 5;707(2):236-42. doi: 10.1016/0167-4838(82)90356-9.

Abstract

Taka-amylase A (1,4-alpha-D-glucan glucanohydrolase, EC 3.2.1.1), which contains a single asparagine-linked oligosaccharide unit, was digested with almond glycopeptidase immobilized on Sepharose 6B at 20 degrees C for 4 h. A maximum of 10% of the parent protein was isolated as apoprotein by column chromatography on Con-A Sepharose. The characteristics of the apoprotein were compared to those of the native Taka-amylase A. The removal of the sugar chain from Taka-amylase. A caused no change in the pH-activity profile or in kinetic parameters of the hydrolysis of soluble starch. The stability of the apoprotein toward changing pH and digestion by proteases did not show any appreciable difference from that of the native Taka-amylase. These results suggest that the carbohydrate moiety of Taka-amylase A is not an essential participant in the catalysis.

摘要

高峰淀粉酶A（1,4-α-D-葡聚糖葡聚糖水解酶，EC 3.2.1.1）含有一个天冬酰胺连接的寡糖单元，在20℃下用固定在琼脂糖6B上的杏仁糖肽酶消化4小时。通过Con-A琼脂糖柱色谱法，最多可分离出10%的亲本蛋白质作为脱辅基蛋白。将脱辅基蛋白的特性与天然高峰淀粉酶A的特性进行了比较。从高峰淀粉酶A上除去糖链，不会改变pH-活性曲线或可溶性淀粉水解的动力学参数。脱辅基蛋白对pH变化的稳定性和蛋白酶消化的稳定性与天然高峰淀粉酶A没有明显差异。这些结果表明，高峰淀粉酶A的碳水化合物部分不是催化作用的必需参与者。