In Vivo Resonance Raman Detection of Ferrous Cytochrome c from Mitochondria of Single Living Yeast Cells
In Vivo Resonance Raman Detection of Ferrous Cytochrome c from Mitochondria of Single Living Yeast Cells
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DOI：
10.1246/cl.2010.270
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发表时间：
2010-02
影响因子：
1.6
通讯作者：
Chikao Onogi;H. Hamaguchi
中科院分区：
文献类型：
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作者：
Chikao Onogi;H. Hamaguchi
活体芽殖酵母细胞（酿酒酵母和巴彦酵母的合子）中线粒体的拉曼光谱已在 532 nm 激发下记录。除了已知的磷脂谱带外，还在 1584、1315、1129、749 和 601 cm -1 处观察到强而尖锐的拉曼谱带。在 632.8 nm 激发下获得的相应线粒体拉曼光谱中未观察到这些拉曼带。它们的峰位置和相对强度与报道的亚铁细胞色素 c 的共振增强拉曼谱带非常吻合，后者在 520 nm 处有 Q 谱带吸收。因此，我们在体内成功检测了亚铁细胞色素 c，它是线粒体电子传递链中的关键中间体之一，无需任何预处理。亚铁细胞色素 c 的体内共振拉曼检测为实时定量追踪单个活细胞线粒体呼吸动力学开辟了新的可能性。
Raman spectra of mitochondria in living budding yeast cells (zygote of Saccharomyces cerevisiae and Saccharomyces bayanus) have been recorded with 532 nm excitation. Strong and sharp Raman bands are observed at 1584, 1315, 1129, 749, and 601 cm -1 , in addition to known phospholipid bands. These Raman bands are not observed in the corresponding Raman spectrum of mitochondria obtained with 632.8 nm excitation. Their peak positions and relative intensities agree excellently with the reported resonance-enhanced Raman bands of ferrous cytochrome c, which has a Q-band absorption at 520 nm. We have thus succeeded in detecting ferrous cytochrome c, one of the key intermediates in the electron-transport chain in mitochondria, in vivo without any pretreatment. In vivo resonance Raman detection of ferrous cytochrome c opens up new possibilities for real-time and quantitative tracing of respiration dynamics in mitochondria in a single living cell.