Quantitative analysis of cohesin complex stoichiometry and SMC3 modification-dependent protein interactions.
Quantitative analysis of cohesin complex stoichiometry and SMC3 modification-dependent protein interactions.
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粘蛋白复合物化学计量和SMC3修饰依赖性蛋白相互作用的定量分析。
DOI：
10.1021/pr2002758
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发表时间：
2011-08-05
影响因子：
4.4
通讯作者：
Qin, Jun
中科院分区：
文献类型：
--
作者：
Ding, Chen;Li, Yehua;Kim, Beom-Jun;Malovannaya, Anna;Jung, Sung Yun;Wang, Yi;Qin, Jun
黏连蛋白是一种蛋白质复合物，在细胞分裂过程中对复制的姐妹染色单体的配对起重要作用。脊椎动物cohesin复合物由四个核心组成部分组成，包括染色体结构维持蛋白SMC 1和SMC 3，RAD 21和SA 2/SA 1。广泛的研究表明，粘着蛋白通过与关闭环的RAD 21蛋白结合的V形SMC 1/SMC 3异二聚体来捕获姐妹染色单体。因此，单“环”模型提出两个姐妹染色单体被困在由4个亚基中的每一个分子组成的单环中。然而，也存在替代模式的证据。手铐模型表明，每个姐妹染色单体被单独捕获的两个环，通过共享的SA1/SA2亚基连接。我们在这里报告的测定内源性粘附素复合物的粘附素亚基的化学计量的定量质谱。使用qConCAT为基础的同位素标记，我们表明，cohesin核心复合物含有等摩尔的4个核心组件，这表明每个cohesin环是由一个SA 1/SA 2分子关闭。此外，我们应用这种策略来量化翻译后修饰依赖的粘附素相互作用。我们证明，定量质谱是一个强大的工具，用于测量内源性蛋白质核心复合物的化学计量。
Cohesin is a protein complex that plays an essential role in pairing replicated sister chromatids during cell division . The vertebrate cohesin complex consists of four core components including structure maintenance of chromosomes proteins SMC1 and SMC3, RAD21 and SA2/SA1. Extensive research suggests that cohesin traps the sister chromatids by a V-shaped SMC1/SMC3 heterodimer bound to the RAD21 protein that closes the ring. Accordingly, the single ‘ring’ model proposes that two sister chromatids are trapped in a single ring that is composed of one molecule each of the 4 subunits. However, evidence also exists for alternative models. The hand-cuff model suggests that each sister chromatid is trapped individually by two rings that are joined through the shared SA1/SA2 subunit. We report here the determination of cohesin subunit stoichiometry of endogenous cohesin complex by quantitative mass spectrometry. Using qConCAT-based isotope labeling, we show that the cohesin core complex contains equimolar of the 4 core components, suggesting that each cohesin ring is closed by one SA1/SA2 molecule. Furthermore, we applied this strategy to quantify post-translational modification-dependent cohesin interactions. We demonstrate that quantitative mass spectrometry is a powerful tool for measuring stoichiometry of endogenous protein core complex.
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影响因子：
7.4
作者：
Jung, Sung Yun;Li, Yehua;Qin, Jun
通讯作者：
Qin, Jun
影响因子：
16
作者：
Haering, CH;Löwe, J;Nasmyth, K
通讯作者：
Nasmyth, K
影响因子：
7
作者：
Rivers, Jenny;Simpson, Deborah M.;Beynon, Robert J.
通讯作者：
Beynon, Robert J.
影响因子：
4.8
作者：
Ong, SE;Foster, LJ;Mann, M
通讯作者：
Mann, M
影响因子：
64.8
作者：
Haering, Christian H.;Farcas, Ana-Maria;Nasmyth, Kim
通讯作者：
Nasmyth, Kim