Noothalapati Hemanth, Shigeto Shinsuke
Department of Applied Chemistry and Institute of Molecular Science, National Chiao Tung University , Hsinchu 30010, Taiwan.
Anal Chem. 2014 Aug 5;86(15):7828-34. doi: 10.1021/ac501735c. Epub 2014 Jul 10.
Understanding cellular metabolism is a major challenge in current systems biology and has triggered extensive metabolomics research, which in most cases involves destructive analysis. However, the information obtainable only in a nondestructive manner will be required for accurately mapping the global structure of the organism's metabolic network at a given instant. Here we report that metabolic pathways can be explored in vivo by mixed stable isotope-labeled Raman microspectroscopy in conjunction with multivariate curve resolution analysis. As a model system, we studied ergosterol biosynthesis in single living fission yeast cells grown in mixtures of normal and (13)C-labeled glucose as the sole carbon source. The multivariate spectral data analysis of space-resolved Raman spectra revealed the intrinsic spectra and relative abundances of all isotopomers of ergosterol whose carbon atoms in the 5,7-diene moiety of the sterol skeleton are either partly or fully substituted with (13)C. Our approach is applicable to other metabolites and will earn a place in the toolbox of metabolomic analysis.
理解细胞代谢是当前系统生物学面临的一项重大挑战，并且引发了广泛的代谢组学研究，而在大多数情况下，代谢组学研究涉及破坏性分析。然而，要在给定时刻准确绘制生物体代谢网络的全局结构，仅以非破坏性方式获取的信息是必需的。在此我们报告，可通过混合稳定同位素标记拉曼光谱结合多元曲线分辨分析在体内探索代谢途径。作为一个模型系统，我们研究了在以正常葡萄糖和（13）C标记葡萄糖的混合物作为唯一碳源生长的单个活裂殖酵母细胞中的麦角固醇生物合成。对空间分辨拉曼光谱的多变量光谱数据分析揭示了麦角固醇所有同位素异构体的本征光谱和相对丰度，这些异构体中固醇骨架5,7 - 二烯部分的碳原子部分或完全被（13）C取代。我们的方法适用于其他代谢物，并将在代谢组学分析工具库中占有一席之地。
