Bioorthogonal chemical reporters for analyzing protein lipidation and lipid trafficking.
Bioorthogonal chemical reporters for analyzing protein lipidation and lipid trafficking.
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DOI：
10.1021/ar200063v
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发表时间：
2011-09-20
影响因子：
18.3
通讯作者：
Charron, Guillaume
中科院分区：
文献类型：
--
作者：
Hang, Howard C.;Wilson, John P.;Charron, Guillaume
蛋白质脂化和脂类运输控制着所有生命王国中的许多关键生物学功能。不同的脂质种类及其与许多蛋白质的共价结合的发现揭示了一个复杂的、受调控的膜和脂化蛋白质网络，这些网络是生理和人类疾病的基本方面的核心。鉴于脂类转运的复杂性和膜脂涉及的蛋白质靶向机制，需要精确和灵敏的方法来监测和鉴定细菌、酵母和高等真核生物中的这些疏水分子。虽然已经开发了许多分析方法来表征膜脂和共价修饰的蛋白质，但传统的试剂和方法灵敏度有限，不能真实地报告感兴趣的脂类，或者不容易获得。生物正交连接反应的发明，如Staudinger连接和叠氮-炔环加成反应，为解决这些局限性提供了新的工具，它们的使用已经开始产生对蛋白质脂化和脂质运输生物学的新见解。在这篇文章中，我们讨论了这些新的生物正交连接反应和脂类化学报告如何为探索脂类修饰蛋白和脂类运输的生物学提供了新的机会。我们实验室和其他几个研究小组的脂类化学记者已经改进了脂肪酰化和预酯化蛋白质的检测和大规模蛋白质组学分析。例如，脂肪酸和类异戊二烯化学报告与生物正交连接方法相结合，避开了放射性类似物的有限敏感性和危害性，使得能够快速而可靠地对所有被测试生物中的脂化蛋白质进行荧光检测。这些化学工具已经揭示了不同细胞状态下蛋白质脂肪作用的变化，并开始为调节机制提供独特的见解。值得注意的是，用脂类化学报告分子标记的蛋白质的纯化使得对脂化蛋白的大规模分析以及涉及新陈代谢、基因表达和先天免疫的新的脂化蛋白的发现成为可能。还开发了特定的脂类记者来监测可溶性脂类的运输；这些物种能够对细胞和组织中的膜进行生物正交成像。未来在生物正交化学、特定的脂类报告和光谱学方面的进展将为脂化蛋白质和膜在生物学中的功能作用提供重要的新见解。
Protein lipidation and lipid trafficking control many key biological functions in all kingdoms of life. The discovery of diverse lipid species and their covalent attachment to many proteins has revealed a complex and regulated network of membranes and lipidated proteins that are central to fundamental aspects of physiology and human disease. Given the complexity of lipid trafficking and the protein targeting mechanisms involved with membrane lipids, precise and sensitive methods are needed to monitor and identify these hydrophobic molecules in bacteria, yeast, and higher eukaryotes. Although many analytical methods have been developed for characterizing membrane lipids and covalently modified proteins, traditional reagents and approaches have limited sensitivity, do not faithfully report on the lipids of interest, or are not readily accessible. The invention of bioorthogonal ligation reactions, such as the Staudinger ligation and azide–alkyne cycloadditions, has provided new tools to address these limitations, and their use has begun to yield fresh insight into the biology of protein lipidation and lipid trafficking. In this Account, we discuss how these new bioorthogonal ligation reactions and lipid chemical reporters afford new opportunities for exploring the biology of lipid-modified proteins and lipid trafficking. Lipid chemical reporters from our laboratory and several other research groups have enabled improved detection and large-scale proteomic analysis of fatty-acylated and prenylated proteins. For example, fatty acid and isoprenoid chemical reporters in conjunction with bioorthogonal ligation methods have circumvented the limited sensitivity and hazards of radioactive analogs, allowing rapid and robust fluorescent detection of lipidated proteins in all organisms tested. These chemical tools have revealed alterations in protein lipidation in different cellular states and are beginning to provide unique insights in mechanisms of regulation. Notably, the purification of proteins labeled with lipid chemical reporters has allowed both the large-scale analysis of lipidated proteins as well as the discovery of new lipidated proteins involved in metabolism, gene expression, and innate immunity. Specific lipid reporters have also been developed to monitor the trafficking of soluble lipids; these species are enabling bioorthogonal imaging of membranes in cells and tissues. Future advances in bioorthogonal chemistry, specific lipid reporters, and spectroscopy should provide important new insight into the functional roles of lipidated proteins and membranes in biology.
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影响因子：
46.2
作者：
Jewett JC;Bertozzi CR
通讯作者：
Bertozzi CR
影响因子：
--
作者：
Charron G;Tsou LK;Maguire W;Yount JS;Hang HC
通讯作者：
Hang HC
影响因子：
64.8
作者：
Ichimura, Y;Kirisako, T;Ohsumi, Y
通讯作者：
Ohsumi, Y
影响因子：
4.9
作者：
Heal, William P.;Wickramasinghe, Sasala R.;Tate, Edward W.
通讯作者：
Tate, Edward W.
DOI：
10.1073/pnas.0907864106
发表时间：
2009-09-08
影响因子：
11.1
作者：
Jao, Cindy Y.;Roth, Mary;Salic, Adrian
通讯作者：
Salic, Adrian