Investigating ROS, RNS, and H2S-Sensitive Signaling Proteins.
Investigating ROS, RNS, and H2S-Sensitive Signaling Proteins.
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DOI：
10.1007/978-1-4939-9463-2_3
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发表时间：
2019
影响因子：
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通讯作者：
Eleanor Williams;M. Whiteman;M. Wood;I. Wilson;M. Ladomery;J. Allainguillaume;T. Teklić;M. Lisjak;J. Hancock
中科院分区：
文献类型：
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作者：
Eleanor Williams;M. Whiteman;M. Wood;I. Wilson;M. Ladomery;J. Allainguillaume;T. Teklić;M. Lisjak;J. Hancock
蛋白质的修饰是改变其活性和功能的关键途径。通常，蛋白质上的巯基、半胱氨酸残基是这种修饰的有吸引力的靶标。假设硫醇基团是可接近的，那么反应可以与细胞中发现的一系列化学物质发生。这些可包括活性氧物质（ROS），例如过氧化氢（H2 O2），活性氮物质，例如一氧化氮（NO）、硫化氢（H2S）或谷胱甘肽。这种修饰通常有助于重要的细胞信号传导过程，其最终导致生物体生理学的修饰。因此，需要能够识别这种修改。有各种各样的技术来寻找蛋白质，可以改变这种方式，但在这里的重点是两种方法：首先，使用荧光巯基衍生物和随后使用质谱法来确定所涉及的硫醇;其次，确认这种变化使用生化测定和遗传突变体。讨论将基于两种模式生物的使用：首先是植物拟南芥（作为细胞培养物和整个植物），其次是线虫线虫秀丽隐杆线虫。然而，如所描述的，这些工具可以用于更广泛的生物系统，包括人类和人类组织培养物。
The modification of proteins is a key way to alter their activity and function. Often thiols, cysteine residues, on proteins are attractive targets for such modification. Assuming that the thiol group is accessible then reactions may take place with a range of chemicals found in cells. These may include reactive oxygen species (ROS), such as hydrogen peroxide (H2O2), reactive nitrogen species such as nitric oxide (NO), hydrogen sulfide (H2S), or glutathione. Such modifications often are instrumental to important cellular signaling processes, which ultimately result in modification of physiology of the organism. Therefore, there is a need to be able to identify such modifications. There are a variety of techniques to find proteins which may be altered in this way but here the focus is on two approaches: firstly, the use of fluorescent thiol derivatives and the subsequent use of mass spectrometry to identify the thiols involved; secondly the confirmation of such changes using biochemical assays and genetic mutants. The discussion will be based on the use of two model organisms: firstly the plantArabidopsis thaliana(both as cell cultures and whole plants) and secondly the nematode wormCaenorhabditis elegans. However, these tools, as described, may be used in a much wider range of biological systems, including human and human tissue cultures.