Ubiquitin Chain Enrichment Middle-Down Mass Spectrometry (UbiChEM-MS) Reveals Cell-Cycle Dependent Formation of Lys11/Lys48 Branched Ubiquitin Chains.
Ubiquitin Chain Enrichment Middle-Down Mass Spectrometry (UbiChEM-MS) Reveals Cell-Cycle Dependent Formation of Lys11/Lys48 Branched Ubiquitin Chains.
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DOI：
10.1021/acs.jproteome.7b00381
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发表时间：
2017-09-01
影响因子：
4.4
通讯作者：
Strieter ER
中科院分区：
文献类型：
--
作者：
Rana ASJB;Ge Y;Strieter ER
细胞信号事件的动力学受到一组不同的泛素链的严格调控。最近的工作表明，分支泛素链组成的Lys 11和Lys 48异肽连接在调节细胞周期进程中发挥着关键作用。然而，不能检测到内源性Lys 11/Lys 48支链。通过将Lys 11连接特异性抗体与高分辨率中-下质谱（称为UbiChEM-MS的方法）相结合，我们试图鉴定细胞中内源性Lys 11/Lys 48分支泛素链。使用异步细胞，我们发现，Lys 11连接的支链只能检测后，与蛋白酶体和非选择性去泛素化酶抑制剂共处理。从有丝分裂停滞释放细胞导致Lys 11/Lys 48支链的显著积累，其中分支点占总泛素群体的~3-4%。该报告强调了UbiChEM-MS在表征各种细胞条件下Lys 11 Ub链结构中的实用性，并证实了有丝分裂过程中Lys 11/Lys 48支链的形成。
The dynamics of cellular signaling events are tightly regulated by a diverse set of ubiquitin chains. Recent work has suggested that branched ubiquitin chains composed of Lys11 and Lys48 isopeptide linkages play a critical role in regulating cell cycle progression. Yet, endogenous Lys11/Lys48 branched chains could not be detected. By combining a Lys11 linkage specific antibody with high-resolution middle-down mass spectrometry (an approach termed UbiChEM-MS) we sought to identify endogenous Lys11/Lys48 branched ubiquitin chains in cells. Using asynchronous cells, we find that Lys11-linked branched chains can only be detected upon cotreatment with a proteasome and nonselective deubiquitinase inhibitor. Releasing cells from mitotic arrest results in a marked accumulation of Lys11/Lys48 branched chains in which branch points represent ~3–4% of the total ubiquitin population. This report highlights the utility of UbiChEM-MS in characterizing the architecture of Lys11 Ub chains under various cellular conditions and corroborates the formation of Lys11/Lys48 branched chains during mitosis.
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影响因子：
44.1
作者：
Swatek KN;Komander D
通讯作者：
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DOI：
10.1007/s13361-013-0789-4
发表时间：
2014-03
影响因子：
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影响因子：
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作者：
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