Neurobiology Sector, SISSA-ISAS, Trieste, Italy.
Biochem Biophys Res Commun. 2010 May 28;396(2):258-64. doi: 10.1016/j.bbrc.2010.04.075. Epub 2010 Apr 23.
Cycloheximide (CHX) is one of the most interesting protein synthesis inhibitors. For this reason, fluorescent derivatives of CHX could find useful applications in cell biology. We report the successful synthesis of a set of novel fluorescent derivatives of CHX. The effect of different functional groups on the biological activity of CHX was studied upon their modification through suitable strategies, i.e., acetylation of the hydroxyl group and reductive amination of the ketone group. The first route induced a complete loss of biological activity, while the second approach allowed a retained inhibition of protein synthesis, as demonstrated by in vitro translation assays. Various fluorescent dyes for reductive amination were tested (i.e., ANTS, APTS, and Rhodamine-123), and the success of the syntheses was demonstrated by diverse analytical techniques. Cycloheximide labeling with fluorescent dyes is a promising approach for developing fluorescence reporters for various applications, both in vitro (fluorescence spectroscopy) and in vivo (live imaging).
环己酰亚胺 (CHX) 是最有趣的蛋白质合成抑制剂之一。出于这个原因，CHX 的荧光衍生物在细胞生物学中可能有有用的应用。我们报告了一组新型 CHX 荧光衍生物的成功合成。通过适当的策略（即羟基的乙酰化和酮基的还原胺化）对 CHX 的不同官能团进行修饰，研究了它们对 CHX 生物活性的影响。第一条途径导致完全丧失生物活性，而第二种方法允许保留对蛋白质合成的抑制作用，这通过体外翻译测定得到证实。测试了用于还原胺化的各种荧光染料（即 ANTS、APTS 和 Rhodamine-123），并通过多种分析技术证明了合成的成功。用荧光染料标记环己酰亚胺是开发各种应用的荧光报告物的有前途的方法，无论是在体外（荧光光谱学）还是在体内（活细胞成像）。
