Variable-Temperature Electrospray Ionization for Temperature-Dependent Folding/Refolding Reactions of Proteins and Ligand Binding.
Variable-Temperature Electrospray Ionization for Temperature-Dependent Folding/Refolding Reactions of Proteins and Ligand Binding.
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用于蛋白质和配体结合的温度依赖性折叠/重折叠反应的变温电喷雾电离。
DOI：
10.1021/acs.analchem.1c00870
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发表时间：
2021-05-11
影响因子：
7.4
通讯作者：
Russell DH
中科院分区：
文献类型：
--
作者：
McCabe JW;Shirzadeh M;Walker TE;Lin CW;Jones BJ;Wysocki VH;Barondeau DP;Clemmer DE;Laganowsky A;Russell DH
蛋白质的稳定性和结构(S)直接与其局部环境或由溶剂、温度、压力和浓度定义的吉布斯自由能景观相耦合。溶液的pH、离子强度、辅因子、化学伴侣和渗透分子会扰乱化学势，并导致结构、稳定性和功能的进一步变化。目前，没有一种单一的分析技术可以在一次测量中监测这些影响。质谱学和离子迁移率-质谱法在蛋白质、蛋白质复合体甚至膜蛋白质复合体的研究中发挥着越来越重要的作用；然而，除了极少数例外，溶液温度对分析物的稳定性和结构的影响还没有得到彻底的研究(S)。在这里，我们描述了一种新的变温电喷雾电离(Vt-ESI)源，它利用热电芯片来冷却和加热静态ESI发射器中包含的溶液。这种设计允许溶液温度从~5°C到98°C变化，在精确控制的温度变化之间平衡时间较短(<；2分钟)。用泛素和Frataxin展示了Vt-ESI-MS和Vt-ESI-离子迁移率-质谱仪研究冷折叠反应和热折叠反应的性能。使用伴侣蛋白GroEL展示了研究依赖温度的配基结合的仪器性能。
Stabilities and structure(s) of proteins are directly coupled to their local environment or Gibbs free energy landscape as defined by solvent, temperature, pressure, and concentration. Solution pH, ionic strength, cofactors, chemical chaperones, and osmolytes perturb the chemical potential and induce further changes in structure, stability, and function. At present, no single analytical technique can monitor these effects in a single measurement. Mass spectrometry and ion mobility-mass spectrometry play increasingly essential roles in studies of proteins, protein complexes, and even membrane protein complexes; however, with few exceptions, the effects of the solution temperature on the stability and structure(s) of analytes have not been thoroughly investigated. Here, we describe a new variable-temperature electrospray ionization (vT-ESI) source that utilizes a thermoelectric chip to cool and heat the solution contained within the static ESI emitter. This design allows for solution temperatures to be varied from ~5 to 98 °C with short equilibration times (<2 min) between precisely controlled temperature changes. The performance of the apparatus for vT-ESI-mass spectrometry and vT-ESI-ion mobility-mass spectrometry studies of cold- and heat-folding reactions is demonstrated using ubiquitin and frataxin. Instrument performance for studies on temperature-dependent ligand binding is shown using the chaperonin GroEL.
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影响因子：
4.4
作者：
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Heck AJR
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影响因子：
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作者：
Fuller DR;Conant CR;El-Baba TJ;Brown CJ;Woodall DW;Russell DH;Clemmer DE
通讯作者：
Clemmer DE
影响因子：
4.9
作者：
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通讯作者：
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