Carlson Erin E, Cravatt Benjamin F
The Skaggs Institute for Chemical Biology, and Department of Cell Biology, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, California 92037, USA.
Nat Methods. 2007 May;4(5):429-35. doi: 10.1038/nmeth1038. Epub 2007 Apr 8.
Chemical probes that target classes of proteins based on shared functional properties have emerged as powerful tools for proteomics. The metabolome rivals, if not surpasses, the proteome in terms of size and complexity, suggesting that efforts to profile metabolites would also benefit from targeted technologies. Here we apply the principle of chemoselective probes to the metabolome, creating a general strategy to tag, enrich and profile large classes of small molecules from biological systems. Key to success was incorporation of a protease-cleavage step to release captured metabolites in a format compatible with liquid chromatography-mass spectrometry (LC-MS) analysis. This technology, termed metabolite enrichment by tagging and proteolytic release (METPR), is applicable to small molecules of any physicochemical class, including polar, labile and low-mass (<100 Da) compounds. We applied METPR to profile changes in the thiol metabolome of human cancer cells treated with the antioxidant N-acetyl-L-cysteine.
基于共享功能特性靶向蛋白质类别的化学探针已成为蛋白质组学的强大工具。代谢组在大小和复杂性方面即便不超过蛋白质组，也与之相当，这表明代谢物谱分析的工作也将受益于靶向技术。在此，我们将化学选择性探针的原理应用于代谢组，创建了一种通用策略，用于标记、富集和分析生物系统中的大类小分子。成功的关键在于加入蛋白酶切割步骤，以与液相色谱 - 质谱（LC - MS）分析兼容的形式释放捕获的代谢物。这项技术称为通过标记和蛋白水解释放进行代谢物富集（METPR），适用于任何物理化学类别的小分子，包括极性、不稳定和低质量（<100 Da）的化合物。我们应用METPR分析了用抗氧化剂N - 乙酰 - L - 半胱氨酸处理的人类癌细胞硫醇代谢组的变化。
