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In Vitro Analysis of E3 Ubiquitin Ligase Function
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对基联血清无化酶的催化和功能的洞察
Sissy Kalayil1,2, Sagar Bhogaraju1,2, Florian Bonn1
1Institute of Biochemistry II, Goethe University Frankfurt - Medical Faculty, University Hospital, Frankfurt am Main, Germany.
Nature
|May 26, 2018
概括
菌 菌
科学领域:
- 分子生物学
- 结构生物学
- 细菌学
背景情况:
- 传统的ubiquitination使用ATP将ubiquitin连接到基底蛋白.
- 致病性肺炎菌通过Side效应器使用NAD+进行氨酸无处不在.
- 通过Side效应器进行血清化的催化机制尚不清楚.
研究的目的:
- 为了阐明SdeA催化核的分子结构.
- 通过SdeA了解氨酸无处置的机制.
- 确定SdeA基质及其在细菌病变中的作用.
主要方法:
- 用X射线结晶学来确定SdeA催化核的结构.
- 生物化学测定以表征酶活性和基质特异性.
- 感染莱吉欧内拉肺突变的研究.
主要成果:
- SdeA催化核的结构显示出单ADP- 基转移酶 (mART) 和基化酶 (PDE) 域.
- 两个不同的催化点介导着两步的氨酸泛化过程.
- 具有靠近血清的疏水性残留物的失序多是首选的SdeA基质.
- SdeA的致病作用取决于基质的无化,而不是无化池的修饰.
结论:
- 这项研究揭示了SdeA介导的血清化结构和机制.
- SdeA使用独特的mART-PDE催化平台进行基基转移.
- 通过SdeA使基质无处不在对于军团菌的发病至关重要.
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