PURIFICATION AND ASSAY OF A 145-KDA PROTEIN (STOP145) WITH MICROTUBULE-STABILIZING AND MOTILITY BEHAVIOR
PURIFICATION AND ASSAY OF A 145-KDA PROTEIN (STOP145) WITH MICROTUBULE-STABILIZING AND MOTILITY BEHAVIOR
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DOI：
10.1073/pnas.83.3.639
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发表时间：
1986-02-01
影响因子：
11.1
通讯作者：
JOB, D
中科院分区：
文献类型：
--
作者：
MARGOLIS, RL;RAUCH, CT;JOB, D
微管在体外分解的能力受到一种称为STOP（仅稳定微管多肽）的蛋白质因子的深刻影响。在这里，我们报告的STOP蛋白的分离，并确认其活性，如预测的那样，高度亚化学计量微管中的微管蛋白。145-kDa的STOP（STOP 145）蛋白的分离已从分离的冷稳定微管的影响，通过两个柱步骤：DEAE离子交换和钙调蛋白亲和柱。为了确认蛋白质的活性，我们已经制备了针对STOP 145的抗体，并使用抗体连接的亲和柱使用该抗体特异性地去除蛋白质和活性。我们的结论是STOP 145蛋白占所观察到的微管在体外稳定。
The capacity of microtubules to disassemble in vitro is profoundly affected by a protein factor designated STOP (stable tubule only polypeptide). Here we report the isolation of STOP protein and confirm that its activity is, as predicted, highly substoichiometric to the tubulin in microtubules. The isolation of the 145-kDa STOP (STOP145) protein has been effected from isolated cold-stable microtubules by two column steps: DEAE ion-exchange and a calmodulin affinity column. To confirm the protein''s activity we have produced an antibody against STOP145 and have used the antibody to specifically remove the protein and the activity using an antibody-linked affinity column. We conclude that the STOP145 protein accounts for the observed in vitro stabilization of microtubules.