Quantitative Characterisation of Low Abundant Yeast Mitochondrial Proteins Reveals Compensation for Haplo-Insufficiency in Different Environments.
Quantitative Characterisation of Low Abundant Yeast Mitochondrial Proteins Reveals Compensation for Haplo-Insufficiency in Different Environments.
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DOI：
10.3390/ijms23158532
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发表时间：
2022-08-01
影响因子：
5.6
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中科院分区：
文献类型：
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低丰度的膜结合蛋白，如转录因子和分子伴侣的定量已被证明是困难的，即使是最复杂的分析技术。在这里，我们利用和优化非侵入性荧光相关光谱（FCS）用于定量低丰度蛋白质，并作为原理的证明，我们选择了两个相互作用的蛋白质参与酵母中线粒体的分裂，Fis 1 p和Mdv 1 p。在酿酒酵母中，Fis 1 p和Mdv 1 p向线粒体的募集对于细胞器的分裂和细胞中功能性线粒体结构的保留是必不可少的。我们使用FCS在单个GFP标记的活酵母细胞中定量纯合子和杂合子细胞中的蛋白质丰度，并研究环境对蛋白质拷贝数、结合/未结合蛋白质状态和迁移动力学的影响。观察到这两种蛋白质主要定位于线粒体结构，Mdv 1 p结合状态在严格的呼吸环境中显着增加。此外，一个补偿机制，控制Fis 1 p丰度删除一个等位基因后，观察到Fis 1 p，但不是在Mdv 1 p，表明差异调节Fis 1 p和Mdv 1 p蛋白表达。
The quantification of low abundant membrane-binding proteins such as transcriptional factors and chaperones has proven difficult, even with the most sophisticated analytical technologies. Here, we exploit and optimise the non-invasive Fluorescence Correlation Spectroscopy (FCS) for the quantitation of low abundance proteins, and as proof of principle, we choose two interacting proteins involved in the fission of mitochondria in yeast, Fis1p and Mdv1p. In Saccharomyces cerevisiae, the recruitment of Fis1p and Mdv1p to mitochondria is essential for the scission of the organelles and the retention of functional mitochondrial structures in the cell. We use FCS in single GFP-labelled live yeast cells to quantify the protein abundance in homozygote and heterozygote cells and to investigate the impact of the environments on protein copy number, bound/unbound protein state and mobility kinetics. Both proteins were observed to localise predominantly at mitochondrial structures, with the Mdv1p bound state increasing significantly in a strictly respiratory environment. Moreover, a compensatory mechanism that controls Fis1p abundance upon deletion of one allele was observed in Fis1p but not in Mdv1p, suggesting differential regulation of Fis1p and Mdv1p protein expression.
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影响因子：
14.9
作者：
Dubreuil B;Sass E;Nadav Y;Heidenreich M;Georgeson JM;Weill U;Duan Y;Meurer M;Schuldiner M;Knop M;Levy ED
通讯作者：
Levy ED
影响因子：
30.8
作者：
Delneri, Daniela;Hoyle, David C.;Oliver, Stephen G.
通讯作者：
Oliver, Stephen G.
影响因子：
4.8
作者：
Bacia, K;Schwille, P
通讯作者：
Schwille, P
影响因子：
3.7
作者：
Guo Q;Koirala S;Perkins EM;McCaffery JM;Shaw JM
通讯作者：
Shaw JM
影响因子：
3.3
作者：
Keskin A;Akdoğan E;Dunn CD
通讯作者：
Dunn CD