A Guide to Native Mass Spectrometry to determine complex interactomes of molecular machines.

A Guide to Native Mass Spectrometry to determine complex interactomes of molecular machines.
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天然质谱指南,以确定分子机的复杂相互作用。

DOI:
10.1111/febs.15281
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发表时间:
2020-06
期刊:
The FEBS journal
影响因子:
--
通讯作者:
Pastore A
Pastore A
中科院分区:
其他
文献类型:
--
作者:
Puglisi R;Boeri Erba E;Pastore A

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天然质谱是生物学中的一种新兴技术,其提供了以高灵敏度和准确度研究非共价结合复合物的可能性。因此,它允许大分子组件的表征,评估其质量和化学计量,并映射相互作用的表面。在这篇综述中,我们讨论了应用本地质谱的动态分子机器的基础上,多个弱相互作用。在这些机器的研究中,关键是要了解在任何时间点形成的各种复合物以及在何种条件下形成。我们专注于铁-硫簇生物发生机的具体例子,因为这是一个动态机器的原型,需要非常具体和苛刻的实验条件,如厌氧性和保留边缘折叠蛋白质的折叠的需要。我们描述的优势,挑战和目前的限制,该技术提供的例子,从我们自己的经验,并提出可能的未来解决方案。天然质谱(MS)可以成功地应用于研究铁硫簇生物成因的相互作用组。在这篇综述中,我们讨论了如何本地MS光谱可以确定所有的物种存在于溶液中,它们的化学计量和它们的相互作用。
Native mass spectrometry is an emerging technique in biology that gives the possibility to study noncovalently bound complexes with high sensitivity and accuracy. It thus allows the characterization of macromolecular assemblies, assessing their mass and stoichiometries and mapping the interacting surfaces. In this review, we discuss the application of native mass spectrometry to dynamic molecular machines based on multiple weak interactions. In the study of these machines, it is crucial to understand which and under which conditions various complexes form at any time point. We focus on the specific example of the iron–sulfur cluster biogenesis machine because this is an archetype of a dynamic machine that requires very specific and demanding experimental conditions, such as anaerobicity and the need of retaining the fold of marginally folded proteins. We describe the advantages, challenges and current limitations of the technique by providing examples from our own experience and suggesting possible future solutions. Native mass spectrometry (MS) can successfully be applied to study the interactome of iron–sulfur cluster biogenesis. In this review, we discuss how native MS spectra can determine all the species present in solution, their stoichiometry and their interactions.
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