Precisely measured protein lifetimes in the mouse brain reveal differences across tissues and subcellular fractions.
Precisely measured protein lifetimes in the mouse brain reveal differences across tissues and subcellular fractions.
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DOI：
10.1038/s41467-018-06519-0
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发表时间：
2018-10-12
影响因子：
16.6
通讯作者：
Rizzoli SO
中科院分区：
文献类型：
--
作者：
Fornasiero EF;Mandad S;Wildhagen H;Alevra M;Rammner B;Keihani S;Opazo F;Urban I;Ischebeck T;Sakib MS;Fard MK;Kirli K;Centeno TP;Vidal RO;Rahman RU;Benito E;Fischer A;Dennerlein S;Rehling P;Feussner I;Bonn S;Simons M;Urlaub H;Rizzoli SO
脑蛋白的周转对生物体的生存至关重要，其扰动与病理有关。然而，蛋白质的寿命很难在体内获得。它们很容易通过在体外用同位素标记的氨基酸喂养细胞来测量，然后进行质谱分析。体内蛋白质至少有两种来源：来自饲料的标记氨基酸和来自已有蛋白质降解的非标记氨基酸。这使得测量变得困难。在这里，我们通过结合小鼠体内同位素标记、质谱分析和数学建模的工作流程严格地解决了这个问题。我们还建立了几个独立的方法来测试和验证结果。这使我们能够精确测量大约3500种大脑蛋白质的寿命。我们的高精度数据提供了大量具有生物学意义的观察结果，包括途径，细胞器，器官或细胞特异性效应，以及极长寿蛋白（ellp）的综合目录。在蛋白质组学水平上，精确测量动物体内的蛋白质周转率在技术上具有挑战性。在这里，Fornasiero和他的同事使用同位素标记与质谱和数学模型来准确地确定老鼠大脑中的蛋白质寿命
The turnover of brain proteins is critical for organism survival, and its perturbations are linked to pathology. Nevertheless, protein lifetimes have been difficult to obtain in vivo. They are readily measured in vitro by feeding cells with isotopically labeled amino acids, followed by mass spectrometry analyses. In vivo proteins are generated from at least two sources: labeled amino acids from the diet, and non-labeled amino acids from the degradation of pre-existing proteins. This renders measurements difficult. Here we solved this problem rigorously with a workflow that combines mouse in vivo isotopic labeling, mass spectrometry, and mathematical modeling. We also established several independent approaches to test and validate the results. This enabled us to measure the accurate lifetimes of ~3500 brain proteins. The high precision of our data provided a large set of biologically significant observations, including pathway-, organelle-, organ-, or cell-specific effects, along with a comprehensive catalog of extremely long-lived proteins (ELLPs). Measuring precise protein turnover rates in animals is technically challenging at the proteomic level. Here, Fornasiero and colleagues use isotopic labeling with mass spectrometry and mathematical modeling to accurately determine protein lifetimes in the mouse brain
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影响因子：
14.9
作者：
UniProt Consortium
通讯作者：
UniProt Consortium
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