標題: | Exploring Metabolic Pathways in Vivo by a Combined Approach of Mixed Stable Isotope-Labeled Raman Microspectroscopy and Multivariate Curve Resolution Analysis |
作者: | Noothalapati, Hemanth Shigeto, Shinsuke 應用化學系 應用化學系分子科學碩博班 Department of Applied Chemistry Institute of Molecular science |
公開日期: | 5-八月-2014 |
摘要: | 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 C-13-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 C-13. Our approach is applicable to other metabolites and will earn a place in the toolbox of metabolomic analysis. |
URI: | http://dx.doi.org/10.1021/ac501735c http://hdl.handle.net/11536/24839 |
ISSN: | 0003-2700 |
DOI: | 10.1021/ac501735c |
期刊: | ANALYTICAL CHEMISTRY |
Volume: | 86 |
Issue: | 15 |
起始頁: | 7828 |
結束頁: | 7834 |
顯示於類別: | 期刊論文 |