Quantitative analysis of chaperone network throughput in budding yeast.
Quantitative analysis of chaperone network throughput in budding yeast.
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DOI：
10.1002/pmic.201200412
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发表时间：
2013-04
期刊：
影响因子：
3.4
通讯作者：
Hubbard, Simon J.
中科院分区：
文献类型：
--
作者：
Brownridge, Philip;Lawless, Craig;Payapilly, Aishwarya B.;Lanthaler, Karin;Holman, Stephen W.;Harman, Victoria M.;Grant, Christopher M.;Beynon, Robert J.;Hubbard, Simon J.
分子伴侣网络调节真核生物基因组中编码的许多蛋白质的折叠和易位，以及对应激的反应。它在萌芽酵母中得到了特别好的描述，其中63个已知的伴侣已被注释，最近的亲和纯化和MS/MS实验在很大程度上有助于表征伴随的伴侣靶标网络。在这项研究中，我们应用我们的QconCAT方法通过SRM MS直接以绝对值(每个细胞的拷贝数)量化一组酵母伴侣蛋白。首先，我们将这些方法与现有的这些酵母蛋白的定量估计进行比较，突出不同方法之间的差异。其次，我们将结果放在伴侣目标网络的背景下，并显示出个体伴侣的丰度与其目标之间的明显关系。这使得我们第一次能够在非应激模式真核生物中描述蛋白质分子通过单个伴侣及其基团在蛋白质组范围内的‘吞吐量’。结果显示了伴侣类的特殊性，它们显示了不同的总体工作负荷、效率和对其目标的亚细胞定位的偏好。交互作用组数据与量化的新颖集成支持通过分子伴侣对蛋白质水平的重新估计。此外，尽管伴侣蛋白以不到40%的注释蛋白质为靶标，但我们发现它们调节大多数蛋白质分子的折叠(∼占细胞总蛋白质通量的62%)，这突显了它们的重要性。
The network of molecular chaperones mediates the folding and translocation of the many proteins encoded in the genome of eukaryotic organisms, as well as a response to stress. It has been particularly well characterised in the budding yeast, Saccharomyces cerevisiae, where 63 known chaperones have been annotated and recent affinity purification and MS/MS experiments have helped characterise the attendant network of chaperone targets to a high degree. In this study, we apply our QconCAT methodology to directly quantify the set of yeast chaperones in absolute terms (copies per cell) via SRM MS. Firstly, we compare these to existing quantitative estimates of these yeast proteins, highlighting differences between approaches. Secondly, we cast the results into the context of the chaperone target network and show a distinct relationship between abundance of individual chaperones and their targets. This allows us to characterise the ‘throughput’ of protein molecules passing through individual chaperones and their groups on a proteome-wide scale in an unstressed model eukaryote for the first time. The results demonstrate specialisations of the chaperone classes, which display different overall workloads, efficiencies and preference for the sub-cellular localisation of their targets. The novel integration of the interactome data with quantification supports re-estimates of the level of protein throughout going through molecular chaperones. Additionally, although chaperones target fewer than 40% of annotated proteins we show that they mediate the folding of the majority of protein molecules (∼62% of the total protein flux in the cell), highlighting their importance.
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影响因子：
14.9
作者：
Güldener U;Münsterkötter M;Oesterheld M;Pagel P;Ruepp A;Mewes HW;Stümpflen V
通讯作者：
Stümpflen V
影响因子：
64.5
作者：
Albanèse, V;Yam, AYW;Frydman, J
通讯作者：
Frydman, J
影响因子：
11.4
作者：
Dekker, Carien;Roe, S. Mark;Willison, Keith R.
通讯作者：
Willison, Keith R.
影响因子：
64.8
作者：
Gavin, AC;Aloy, P;Superti-Furga, G
通讯作者：
Superti-Furga, G
影响因子：
3.3
作者：
Lawless, Craig;Hubbard, Simon J.
通讯作者：
Hubbard, Simon J.