Mass spectrometry-based direct detection of multiple types of protein thiol modifications in pancreatic beta cells under endoplasmic reticulum stress.
Mass spectrometry-based direct detection of multiple types of protein thiol modifications in pancreatic beta cells under endoplasmic reticulum stress.
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DOI：
10.1016/j.redox.2021.102111
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发表时间：
2021-10
期刊：
影响因子：
11.4
通讯作者：
Qian WJ
中科院分区：
文献类型：
--
作者：
Li X;Day NJ;Feng S;Gaffrey MJ;Lin TD;Paurus VL;Monroe ME;Moore RJ;Yang B;Xian M;Qian WJ
基于硫醇的翻译后修饰(PTM)在氧化还原依赖的调节和信号转导中起着关键作用。功能性半胱氨酸(Cys)作为氧化还原开关，通过多种类型的PTM进行调节。在这里，我们的目标是通过建立直接检测工作流程，在蛋白质组水平上表征硫醇PTMS的复杂性。基于LC-MS/MS的工作流程允许同时定量蛋白质丰度和多种类型的硫醇PTMS。为了验证其有效性，将该工作流程应用于thapsigargin处理的小鼠胰腺β细胞(β-TC-6)，以诱导内质网(ER)应激。这导致了>；9000蛋白质和多种类型的硫醇PTMS的定量，包括肽内二硫化(S-S)、S-谷胱甘肽基化(SSG)、S-亚磺化(SO2H)、S-磺酰化(SO3H)、S-过硫化(SSSH)和S-三硫化(SSSH)。观察到丰度显著变化的蛋白质参与了自噬、未折叠蛋白反应、蛋白泛素化途径和eIF2信号转导等典型途径。此外，在一种或多种类型的PTMS中，观察到约500个Cys位点，其中SSH和S-S是主要的修饰类型。在许多情况下，不同的PTM在不同的酶及其活性部位上的水平都发生了显著的变化，而它们的蛋白丰度却变化不大。这些结果为酶活性的翻译和翻译后独立调控提供了证据。在相同的半胱氨酸残基上观察到的硫醇修饰的复杂性说明了在表征和解释蛋白质硫醇修饰及其功能调节方面的挑战。同时定量蛋白质丰度和多种类型的硫醇PTMS。在同一个Cys位点上观察到多种类型的PtM，用于氧化还原调节蛋白。数据揭示了硫醇PTMS及其调控的复杂性。内质网应激下β细胞翻译和翻译后的独特调控。
Thiol-based post-translational modifications (PTMs) play a key role in redox-dependent regulation and signaling. Functional cysteine (Cys) sites serve as redox switches, regulated through multiple types of PTMs. Herein, we aim to characterize the complexity of thiol PTMs at the proteome level through the establishment of a direct detection workflow. The LC-MS/MS based workflow allows for simultaneous quantification of protein abundances and multiple types of thiol PTMs. To demonstrate its utility, the workflow was applied to mouse pancreatic β-cells (β-TC-6) treated with thapsigargin to induce endoplasmic reticulum (ER) stress. This resulted in the quantification of >9000 proteins and multiple types of thiol PTMs, including intra-peptide disulfide (S–S), S-glutathionylation (SSG), S-sulfinylation (SO2H), S-sulfonylation (SO3H), S-persulfidation (SSH), and S-trisulfidation (SSSH). Proteins with significant changes in abundance were observed to be involved in canonical pathways such as autophagy, unfolded protein response, protein ubiquitination pathway, and EIF2 signaling. Moreover, ~500 Cys sites were observed with one or multiple types of PTMs with SSH and S–S as the predominant types of modifications. In many cases, significant changes in the levels of different PTMs were observed on various enzymes and their active sites, while their protein abundance exhibited little change. These results provide evidence of independent translational and post-translational regulation of enzyme activity. The observed complexity of thiol modifications on the same Cys residues illustrates the challenge in the characterization and interpretation of protein thiol modifications and their functional regulation. Simultaneous quantification of protein abundances and multiple types of thiol PTMs. Multiple types PTMs observed on the same Cys sites for redox-regulated proteins. Data revealed complexity of thiol PTMs and their regulation. Distinctive translational and post-translational regulation under ER stress in β-cells.
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影响因子：
5.5
作者：
Balta E;Kramer J;Samstag Y
通讯作者：
Samstag Y
DOI：
10.3390/antiox10030499
发表时间：
2021-03-23
期刊：
Antioxidants (Basel, Switzerland)
影响因子：
--
作者：
Day NJ;Gaffrey MJ;Qian WJ
通讯作者：
Qian WJ
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影响因子：
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作者：
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影响因子：
2.9
作者：
Majmudar, Jaimeen D.;Martin, Brent R.
通讯作者：
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影响因子：
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作者：
Duan J;Kodali VK;Gaffrey MJ;Guo J;Chu RK;Camp DG;Smith RD;Thrall BD;Qian WJ
通讯作者：
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