Quantitative Structural Proteomics Unveils the Conformational Changes of Proteins under the Endoplasmic Reticulum Stress.
Quantitative Structural Proteomics Unveils the Conformational Changes of Proteins under the Endoplasmic Reticulum Stress.
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DOI：
10.1021/acs.analchem.2c03076
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发表时间：
2022-09-27
影响因子：
7.4
通讯作者：
Wu, Ronghu
中科院分区：
文献类型：
--
作者：
Yin, Kejun;Tong, Ming;Sun, Fangxu;Wu, Ronghu
蛋白质结构决定了它们的活性和与其他分子的相互作用。蛋白质结构和构象变化的全局分析不能通过常用的基于丰度的蛋白质组学来实现。在这里，我们整合了半胱氨酸共价标记，选择性富集和定量蛋白质组学研究蛋白质结构和结构变化的大规模。该方法被应用于在HEK 293 T细胞中的蛋白质结构和在衣霉素（Tm）诱导的内质网（ER）应激的细胞中的蛋白质结构变化的全局研究。我们定量了数千个半胱氨酸残基，这些残基包含了前所未有的和有价值的蛋白质结构信息。将该方法与细胞培养中的氨基酸脉冲稳定同位素标记相结合，我们进一步分析了Tm处理下细胞中预先存在的和新合成的蛋白质之间的折叠状态差异。除了新合成的蛋白质外，出乎意料的是，许多预先存在的蛋白质被发现在ER应激时变得解折叠，特别是那些与基因转录和蛋白质翻译相关的蛋白质。此外，目前的研究结果表明，N-糖基化在新合成的蛋白质的三级和四级结构的折叠过程中起着比二级结构更重要的作用。考虑到半胱氨酸在蛋白质结构中的重要性，该方法可广泛应用于生物和生物医学研究领域。
Protein structures are decisive for their activities and interactions with other molecules. Global analysis of protein structures and conformational changes cannot be achieved by commonly used abundance-based proteomics. Here, we integrated cysteine covalent labeling, selective enrichment, and quantitative proteomics to study protein structures and structural changes on a large scale. This method was applied to globally investigate protein structures in HEK293T cells and protein structural changes in the cells with the tunicamycin (Tm)-induced endoplasmic reticulum (ER) stress. We quantified several thousand cysteine residues, which contain unprecedented and valuable information of protein structures. Combining this method with pulsed stable isotope labeling by amino acids in cell culture, we further analyzed the folding state differences between pre-existing and newly synthesized proteins in cells under the Tm treatment. Besides newly synthesized proteins, unexpectedly, many pre-existing proteins were found to become unfolded upon ER stress, especially those related to gene transcription and protein translation. Furthermore, the current results reveal that N-glycosylation plays a more important role in the folding process of the tertiary and quaternary structures than the secondary structures for newly synthesized proteins. Considering the importance of cysteine in protein structures, this method can be extensively applied in the biological and biomedical research fields.
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影响因子：
64.5
作者：
Huttlin EL;Jedrychowski MP;Elias JE;Goswami T;Rad R;Beausoleil SA;Villén J;Haas W;Sowa ME;Gygi SP
通讯作者：
Gygi SP
影响因子：
62.1
作者：
Liu XR;Zhang MM;Gross ML
通讯作者：
Gross ML
DOI：
10.1073/pnas.96.22.12494
发表时间：
1999-10-26
影响因子：
11.1
作者：
Baud, F;Karlin, S
通讯作者：
Karlin, S
影响因子：
4.4
作者：
Kudelko, Mateusz;Chan, Cecilia W. L.;Chan, Danny
通讯作者：
Chan, Danny
聚糖的疏水性和糖基化位点如何影响蛋白质折叠和稳定性：分子动力学模拟
影响因子：
3.3
作者：
Lu, Diannan;Yang, Cheng;Liu, Zheng
通讯作者：
Liu, Zheng