Photobleaching of the "Raman Spectroscopic Signature of Life" and Mitochondrial Activity in Rho- Budding Yeast Cells
Photobleaching of the "Raman Spectroscopic Signature of Life" and Mitochondrial Activity in Rho- Budding Yeast Cells
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DOI：
10.1021/jp903478r
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发表时间：
2009-08-06
影响因子：
3.3
通讯作者：
Hamaguchi, Hiro-O
中科院分区：
文献类型：
--
作者：
Onogi, Chikao;Hamaguchi, Hiro-O
对两种类型的四倍体酵母野生型菌株和Rho(-)菌株的线粒体拉曼光谱进行了测量和比较。在两株强度相似的菌株中都发现了“LIFE的拉曼光谱特征”，即在1602厘米(-1)处有一个强烈的拉曼谱带，敏锐地反映了线粒体的活性。这一特征显示了在632.8 nm He-Ne激光照射下的光漂白，仅对Rho(-)菌株。在没有激光照射的情况下，将细胞置于黑暗中两小时，光漂白就会恢复。这些发现表明，产生1602 cm(-1)带的分子物种是在代谢周期中形成和积累的中间产物，它在632.8 nm处被吸收进行光解。通过对时间分辨拉曼光谱的SVD分析，我们还在400-1200 cm(-1)区域发现了一些与1602 cm(-1)谱带具有相同时间行为的弱拉曼光谱，从而获得了一些关于“生命的拉曼光谱特征”起源的新信息。
Raman spectra of mitochondria in single living budding yeast (zygote of Saccharomyces cerevisiae and Saccharomyces bayanus) cells have been measured and compared for two types of tetraploid strains, the wild-type strain and the rho(-) strain. The "Raman spectroscopic signature of life", a strong Raman band at 1602 cm(-1) that sharply reflects the mitochondrial activity, has been found in both of the two strains with similar intensities. This signature showed photobleaching under irradiation of the 632.8 nm line of a He-Ne laser, only for the rho(-) strain. The photobleaching recovered by keeping the cell in the dark for two hours without laser irradiation. These findings indicate that the molecular species that gives rise to the 1602 cm(-1) band is an intermediate formed and accumulated during the metabolic cycle and that it absorbs at 632.8 nm to undergo photodecomposition. We also found a number of weak Raman bands in the 400-1200 cm(-1) region that show the same temporal behaviors as that of the 1602 cm(-1) band by an SVD analysis of time-resolved Raman spectra, Several new pieces of information oil the origin of the "Raman spectroscopic signature of life" have thus been obtained.