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Assaying Protein Kinase Activity with Radiolabeled ATP
Published on: May 26, 2017
控制由基因诱导的氨酸/氨酸酸酶PAC1控制MAP激酶激活
1Laboratory of Pathology, National Cancer Institute, Bethesda, Maryland 20892.
Nature
|February 17, 1994
概括
PAC1是一种酸酶,可以使基激活蛋白 (MAP) 激酶失活. 这一发现揭示了PAC1的存在.
科学领域:
- 蜂信号传输是如何进行的
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 中原刺激激活了涉及中原激活蛋白 (MAP) 激酶ERK1和ERK2.2的细胞内信号级联.
- MAP 激酶激活是通过 MAP 激酶激酶 (MEK) 的双酸化发生的.
- 已激活的MAP激酶转移到核中,酸化转录因子.
研究的目的:
- 为了研究PAC1的作用,之前确定的核氨酸酸酶,在MAP激酶调节中.
- 为了确定PAC1是否作为一个生理学上相关的MAP激酶酸酶.
主要方法:
- 在实验室中将重组PAC1描述为双特异性酸酶.
- 评估PAC1对MAP激酶的基质特异性.
- 涉及构成性PAC1表达的体内研究,以评估其对MAP激酶活性的影响.
- 在PAC1表达后对MAP激酶调节的记者基因表达的分析.
主要成果:
- 再组合的PAC1显示了双特异的氨酸/氨酸酸酶活性,对MAP激酶具有很高的特异性.
- 构成性PAC1表达抑制了由表皮生长因子,博乙酸和T细胞受体交联刺激的MAP激酶活性.
- 通过PAC1介导的MAP激酶的失活导致了MAP激酶调节的记者基因表达的抑制.
结论:
- PAC1是一种生理相关的MAP激酶酸酶.
- PAC1在调节MAP激酶信号通路方面发挥着至关重要的作用.
- MAP 激酶的 PAC1 失活影响下游的转录调节.
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