Exploring an Alternative Cysteine-Reactive Chemistry to Enable Proteome-Wide PPI Analysis by Cross-Linking Mass Spectrometry.
Exploring an Alternative Cysteine-Reactive Chemistry to Enable Proteome-Wide PPI Analysis by Cross-Linking Mass Spectrometry.
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DOI：
10.1021/acs.analchem.2c04986
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发表时间：
2023-01-31
影响因子：
7.4
通讯作者：
Huang, Lan
中科院分区：
文献类型：
--
作者：
Jiao, Fenglong;Salituro, Leah J.;Yu, Clinton;Gutierrez, Craig B.;Rychnovsky, Scott D.;Huang, Lan
MS可裂解交联质谱(XL-MS)的发展使得无需细胞工程就能在全球范围内有效地捕捉和鉴定内源蛋白质-蛋白质相互作用(PPI)及其残基接触。到目前为止，只有赖氨酸反应性交联剂被成功地应用于蛋白质组范围的PPI谱分析。然而，赖氨酸交联剂本身并不能揭示细胞中完整的PPI图。以前，我们已经开发了一种基于马来酰亚胺的半胱氨酸反应性MS可裂解交联剂(双马来酰亚胺亚硫醚(BMSO))，它可以有效地定位蛋白质复合体的PPI，以产生与赖氨酸反应性试剂互补的相互作用接触。虽然成功了，但马来酰亚胺在生理pH下的水解性和有限的选择性使得它们在蛋白质组范围内的XL-MS中的应用具有挑战性。为了实现全球PPI定位，我们探索了一种替代的半胱氨酸标记化学，从而设计并合成了一种含亚砜的MS可裂解卤代乙酰胺交联剂DBrASO(DBrASO)。我们的结果表明，DBrASO交联肽表现出与其他含亚砜的MS可裂解交联剂相同的裂解特征，从而使它们能够被MSN明确识别。结合新开发的二维肽分离方法，我们成功地对HEK293细胞裂解产物进行了基于DBrASO的XL-MS分析，并证明了其在系统水平上补充赖氨酸反应性试剂和扩大PPI覆盖范围的能力。
The development of MS-cleavable cross-linking mass spectrometry (XL-MS) has enabled effective capture and identification of endogenous protein-protein interactions (PPIs) and their residue contacts at the global scale without cell engineering. So far, only lysine-reactive cross-linkers have been successfully applied for proteome-wide PPI profiling. However, lysine cross-linkers alone cannot uncover the complete PPI map in cells. Previously we have developed a maleimide-based cysteine-reactive MS-cleavable cross-linker (bismaleimide sulfoxide(BMSO)) that is effective for mapping PPIs of protein complexes to yield interaction contacts complementary to lysine-reactive reagents. While successful, the hydrolysis and limited selectivity of maleimides at physiological pH make their applications in proteome-wide XL-MS challenging. To enable global PPI mapping, we have explored an alternative cysteine labeling chemistry, and thus designed and synthesized a sulfoxide-containing MS-cleavable haloacetamide-based cross-linker, Dibromoacetamide sulfOxide (DBrASO). Our results have demonstrated that DBrASO cross-linked peptides display the same fragmentation characteristics as other sulfoxide-containing MS-cleavable cross-linkers, permitting their unambiguous identification by MSn. In combination with a newly developed two-dimensional peptide fractionation method, we have successfully performed DBrASO-based XL-MS analysis of HEK293 cell lysates and demonstrated its capability to complement lysine-reactive reagents and expand PPI coverage at the systems-level.
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影响因子：
16.8
作者：
Munoz, Ines G.;Yebenes, Hugo;Montoya, Guillermo
通讯作者：
Montoya, Guillermo
DOI：
10.1073/pnas.1915542117
发表时间：
2020-02-25
影响因子：
11.1
作者：
Gutierrez, Craig;Chemmama, Ilan E.;Huang, Lan
通讯作者：
Huang, Lan
影响因子：
56.9
作者：
Herzog, Franz;Kahraman, Abdullah;Aebersold, Ruedi
通讯作者：
Aebersold, Ruedi
影响因子：
7.4
作者：
Gutierrez, Craig B.;Block, Sarah A.;Huang, Lan
通讯作者：
Huang, Lan
影响因子：
7
作者：
Mueller, Torsten;Winter, Dominic
通讯作者：
Winter, Dominic