Evidence that the tightly bound magnesium in tubulin is associated with the N-site GTP.
Evidence that the tightly bound magnesium in tubulin is associated with the N-site GTP.
复制标题
有证据表明微管蛋白中紧密结合的镁与 N 位 GTP 相关。
DOI：
10.1016/0014-5793(90)80513-i
复制
发表时间：
1990
期刊：
影响因子：
3.5
通讯作者：
Himes,RH
中科院分区：
文献类型：
--
作者：
Osei,AA;Everett,GW;Himes,RH
为了确定在纯化微管蛋白中发现的紧密结合的Mg2+是否与N位GTP或E位GDP或GTP相关，我们通过几种方法去除E位核苷酸：(i)碱性磷酸酶处理；（ii）使用过量的GMPPCP置换；（iii）在碱性磷酸酶和不可水解类似物存在的情况下聚合微管蛋白。在不发生变性的条件下，Mg2+的含量保持在1微管蛋白左右。此外，Mg/GTP比值始终保持为1。这些结果表明Mg2+与N位GTP相关。
In an attempt to determine whether the tightly bound Mg2+found in purified tubulin in associated with the N‐site GTP or the E‐site GDP or GTP, we removed the E‐site nucleotide by several means: (i) alkaline phosphatase treatment; (ii) displacement using excess GMPPCP; and (iii) polymerizing tubulin in the presence of alkaline phosphatase and non‐hydrolyzable analogues. The Mg2+content remained equal to about 1 tubulin under conditions where denaturation did not occur. Moreover, the Mg/GTP ratio always remained equal to 1. These results indicate that the Mg2+is associated with the N‐site GTP.