Online protein unfolding characterized by ion mobility electron capture dissociation mass spectrometry: cytochrome C from neutral and acidic solutions
Online protein unfolding characterized by ion mobility electron capture dissociation mass spectrometry: cytochrome C from neutral and acidic solutions
复制标题
以离子淌度电子捕获解离质谱为特征的在线蛋白质展开：中性和酸性溶液中的细胞色素 C
DOI：
10.1007/s00216-022-04501-w
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发表时间：
2023
影响因子：
4.3
通讯作者：
Webb, Ian K.
中科院分区：
文献类型：
--
作者：
Cain, Rebecca L.;Webb, Ian K.
电喷雾离子化质谱（ESI-MS）实验，包括离子迁移谱质谱（ESI-IMS-MS）和电子捕获解离（ECD）的蛋白质从水溶液中电离，已被用于研究完整蛋白质的溶液样结构。通过在ESI之前在线混合水性蛋白质与变性剂，可以精确控制和快速分析蛋白质解折叠的量，从而可以表征蛋白质折叠途径中的蛋白质折叠中间体。在此，我们将各种pH值的溶液与含水细胞色素C在线混合，用于展开，并用ESI-MS电荷态分布测量，IMS和ECD表征其展开中间体。折叠中间体和未折叠的细胞色素c结构的存在下，检测到的电荷状态，到达时间分布（ATDs），和ECD的变化。我们还比较了在“温和”（即，低能量）离子传输条件下测量的结构（即，更高的能量）传输。这项工作证实，当使用“温和”的仪器条件下，气相细胞色素c离子反映的各种溶液相结构的属性。然而，最大化离子传输的“苛刻”条件会产生不再与溶液结构变化相关的扩展结构。图形摘要
Electrospray ionization mass spectrometry (ESI-MS) experiments, including ion mobility spectrometry mass spectrometry (ESI-IMS-MS) and electron capture dissociation (ECD) of proteins ionized from aqueous solutions, have been used for the study of solution-like structures of intact proteins. By mixing aqueous proteins with denaturants online before ESI, the amount of protein unfolding can be precisely controlled and rapidly analyzed, permitting the characterization of protein folding intermediates in protein folding pathways. Herein, we mixed various pH solutions online with aqueous cytochrome C for unfolding and characterizing its unfolding intermediates with ESI-MS charge state distribution measurements, IMS, and ECD. The presence of folding intermediates and unfolded cytochrome c structures were detected from changes in charge states, arrival time distributions (ATDs), and ECD. We also compared structures from nondenaturing and denaturing solution mixtures measured under “gentle” (i.e., low energy) ion transmission conditions with structures measured under “harsh” (i.e., higher energy) transmission. This work confirms that when using “gentle” instrument conditions, the gas-phase cytochrome c ions reflect attributes of the various solution-phase structures. However, “harsh” conditions that maximize ion transmission produce extended structures that no longer correlate with changes in solution structure.Graphical abstract
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DOI：
10.1111/j.1742-4658.2011.08376.x
发表时间：
2011-11
期刊：
The FEBS journal
影响因子：
--
作者：
Stevens JM;Mavridou DA;Hamer R;Kritsiligkou P;Goddard AD;Ferguson SJ
通讯作者：
Ferguson SJ
影响因子：
2.9
作者：
GOTO, Y;HAGIHARA, Y;NISHII, I
通讯作者：
NISHII, I
探讨多相氧化修饰对溶液和气相中细胞色素 c 稳定性的影响。
影响因子：
3.2
作者：
Victor C Yin;L. Konermann
通讯作者：
L. Konermann
影响因子：
15
作者：
Breuker, K;Oh, HB;McLafferty, FW
通讯作者：
McLafferty, FW
影响因子：
7.8
作者：
Chavez JD;Bruce JE
通讯作者：
Bruce JE