Gopalakrishnan T V, Thompson E B
J Biol Chem. 1977 Apr 25;252(8):2717-25.
Concanavalin A added to intact cells at 37 degrees caused rapid and reversible inactivation of a soluble enzyme, tyrosine aminotransferase, in two lines of rat hepatoma tissue culture cells grown in monolayer culture. This temperature-dependent process was independent of de novo protein and RNA synthesis and independent of increased uptake of Ca2+ and Mg2+ or glucose. The inactivation could be reversed by adding alpha-methyl-D-mannopyranoside a competing sugar for concanavalin A binding. Other lectins known to bind to different sugars did not bring about the inactivation of tyrosine aminotransferase. Addition of concanavalin A did not result in the inactivation of another soluble enzyme, lactic dehydrogenase. The maintenance of tyrosine aminotransferase in an inactive form after the binding of concanavalin A to the cells required the continued presence of concanavalin A. This effect of concanavalin A could not be mimicked either by dibutyryl cyclic adenosine or guanosine monophosphoric acid. Incubation of cell extracts with concanavalin A did not result in inactivation nor did mixing of extracts from concanavalin A-treated cells with extracts from untreated cells. On the basis of these results we conclude that the following are the essential requirements for concanavalin A to bring about the inactivation of tyrosine aminotransferase: (a) the binding of native concanavalin A to the cells; (b) integrity of certain structural elements of the cells.
在37摄氏度下向完整细胞中添加伴刀豆球蛋白A,可使单层培养的两株大鼠肝癌组织培养细胞中的一种可溶性酶——酪氨酸转氨酶迅速且可逆地失活。这个温度依赖性过程与从头合成蛋白质和RNA无关,也与增加对Ca2+、Mg2+或葡萄糖的摄取无关。通过添加α-甲基-D-甘露吡喃糖苷(一种与伴刀豆球蛋白A结合的竞争性糖类)可使失活逆转。已知能与不同糖类结合的其他凝集素不会导致酪氨酸转氨酶失活。添加伴刀豆球蛋白A不会导致另一种可溶性酶——乳酸脱氢酶失活。伴刀豆球蛋白A与细胞结合后,酪氨酸转氨酶维持无活性形式需要伴刀豆球蛋白A持续存在。伴刀豆球蛋白A的这种作用既不能被二丁酰环腺苷或鸟苷单磷酸模拟。用伴刀豆球蛋白A孵育细胞提取物不会导致失活,将伴刀豆球蛋白A处理过的细胞提取物与未处理细胞的提取物混合也不会导致失活。基于这些结果,我们得出结论,伴刀豆球蛋白A导致酪氨酸转氨酶失活的基本要求如下:(a)天然伴刀豆球蛋白A与细胞结合;(b)细胞某些结构元件的完整性。