Probing mitochondrial metabolism in vivo
Probing mitochondrial metabolism in vivo
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DOI：
10.1073/pnas.1819614116
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发表时间：
2018-12
期刊：
影响因子：
--
通讯作者：
Gregory S. McElroy;N. Chandel
中科院分区：
文献类型：
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作者：
Gregory S. McElroy;N. Chandel
线粒体代谢包括产生ATP以驱动细胞内不利的能量反应并产生大分子合成所需的构建单元的途径。在过去的二十年中，已经清楚的是，线粒体代谢并不作为生物学的潜在机制发挥被动作用，而是通过释放活性氧和代谢物来控制基因表达，从而在决定细胞命运和功能方面发挥动态和有影响力的作用（1）。线粒体酶和复合物的区室化对于维持细胞内的重要信号通路是必不可少的。因此，线粒体基质和细胞质之间代谢物的特性和浓度显著不同。迄今为止，我们对线粒体代谢物浓度的大部分理解仅限于体外环境（2）。在PNAS中，Bayraktar等人（3）建立了MITO-Tag小鼠，这是一种通过快速评估线粒体代谢物对了解体内线粒体代谢产生重大影响的工具。MITO-Tag小鼠利用线粒体外膜蛋白OMP 25的线粒体定位序列将含有来自流感血凝素和GFP的HA表位的融合蛋白靶向线粒体。该MITO-Tag先前已发表用于体外使用（2，4）。这种方法允许使用HA表位特异性抗体从体外培养的细胞中快速分离标记的线粒体。现在，在PNAS中，Bayraktar等人（3）通过使用Cre-Lox重组技术在体内特定细胞类型中产生表达MITO-Tag的小鼠模型（图1）采取了重要的下一步。Fig. 1. MITO-Tag小鼠使用Cre-Lox重组在特定细胞类型或组织中表达靶向表位标签。这种表位标签允许从线粒体中快速和直接分离线粒体[1][2] 1。电子邮件：nav{at}northwestern.edu. [1]：#xref-corresp-1-1
Mitochondrial metabolism encompasses pathways that generate ATP to drive intracellular unfavorable energetic reactions and produce the building blocks necessary for macromolecule synthesis. In the past two decades, it has become clear that mitochondrial metabolism does not play a passive role as an underlying mechanism of biology, but instead plays a dynamic and influential role in dictating cell fate and function by controlling gene expression through release of reactive oxygen species and metabolites (1). The compartmentalization of mitochondrial enzymes and complexes is essential for the maintenance of important signaling pathways within the cell. For this reason, the identity and concentration of metabolites is substantially different between the mitochondrial matrix and the cytosol. To date, much of our understanding of the concentration of mitochondrial metabolites has been limited to in vitro settings (2). In PNAS, Bayraktar et al. (3) establish MITO-Tag Mice, a tool that will have a substantial impact on the understanding of in vivo mitochondrial metabolism by allowing rapid assessment of mitochondrial metabolites. MITO-Tag Mice take advantage of the mitochondrial localization sequences of the outer mitochondrial membrane protein OMP25 to target a fusion protein containing the HA epitope from influenza hemagglutinin and GFP to the mitochondria. This MITO-Tag was previously published for in vitro use (2, 4). This methodology allowed the rapid isolation of tagged mitochondria from cells cultured in vitro using antibodies specific to the HA epitope. Now, in PNAS, Bayraktar et al. (3) take the important next step by generating a mouse model that expresses the MITO-Tag in specific cell types in vivo, using Cre-Lox recombination technology (Fig. 1). Fig. 1. MITO-Tag Mice use Cre-Lox recombination to express a mitochondria-targeted epitope tag in specific cell types or tissues. This epitope tag allows the rapid and direct isolation of mitochondria from the … [↵][1]1To whom correspondence should be addressed. Email: nav{at}northwestern.edu. [1]: #xref-corresp-1-1