Monitoring copopulated conformational states during protein folding events using electrospray ionization-ion mobility spectrometry-mass spectrometry.
Monitoring copopulated conformational states during protein folding events using electrospray ionization-ion mobility spectrometry-mass spectrometry.
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DOI：
10.1016/j.jasms.2007.09.017
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发表时间：
2007-12
影响因子：
3.2
通讯作者：
Ashcroft AE
中科院分区：
文献类型：
--
作者：
Smith DP;Giles K;Bateman RH;Radford SE;Ashcroft AE
蛋白质折叠的确切机制仍然难以捉摸，而且缺乏能够监测折叠过程中共生蛋白质构象个体行为的生物物理技术。本文采用离子迁移率谱(IMS)和电喷雾电离质谱仪(ESI-MS)相结合的方法，成功地分离和分析了不同截面和/或不同电荷状态的蛋白质构象。在最初的测试中，分离出了蛋白质细胞色素c的折叠和部分折叠构象的集合。对淀粉样蛋白β2-微球蛋白(β2M)进行的一项详细研究对其折叠情况产生了重要的见解，该蛋白通过蛋白质展开然后自我聚集形成纤维，并导致疾病透析相关的淀粉样变性。最初，使用电喷雾离子质谱仪在pH 2至7的范围内系统滴定β2M，允许在整个酸变性过程中监测和定量单个构象。此外，野生型β2M与具有一系列折叠稳定性和淀粉样纤维形成倾向的单氨基酸和双氨基酸变体的比较，为不同构象在蛋白质稳定性和淀粉样变性聚集中的作用提供了具有启发性的证据。本文提供的ESI-IMS-MS数据不仅展示了ESI-MS的一个重要和信息丰富的进一步维度，而且还说明了ESI-IMS-MS技术在揭开蛋白质折叠之谜以及特别是研究蛋白质错折叠疾病方面的潜力。
The precise mechanism of protein folding remains elusive and there is a deficiency of biophysical techniques that are capable of monitoring the individual behavior of copopulated protein conformers during the folding process. Herein, an ion mobility spectrometry (IMS) device integrated with electrospray ionization mass spectrometry (ESI-MS) has been used to successfully separate and analyze protein conformers differing in cross section and/or charge state. In an initial test, an ensemble of folded and partially folded conformers of the protein cytochrome c was separated. A detailed study undertaken on the amyloidogenic protein β2-microglobulin (β2m), which forms fibrils by protein unfolding followed by self-aggregation and is responsible for the disease dialysis-related amyloidosis, has generated important insights into its folding landscape. Initially, a systematic titration of β2m over the pH range 2 to 7 using ESI-IMS-MS allowed individual conformers to be monitored and quantified throughout the acid denaturation process. Furthermore, a comparison of wild-type β2m with single and double amino acid variants with a range of folding stabilities and propensities for amyloid fibril formation has provided illuminating evidence of the role of different conformers in protein stability and amyloidogenic aggregation. The ESI-IMS-MS data presented here not only demonstrate an important and informative further dimension to ESI-MS, but also illustrate the potential of the ESI-IMS-MS technique for unravelling protein folding enigmas in general and studying protein misfolding diseases in particular.
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影响因子：
2.9
作者：
GOTO, Y;HAGIHARA, Y;NISHII, I
通讯作者：
NISHII, I
影响因子：
7.4
作者：
Badman, ER;Hoaglund-Hyzer, CS;Clemmer, DE
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DOI：
10.1016/1044-0305(93)85025-s
发表时间：
1993-08-01
影响因子：
3.2
作者：
COVEY, T;DOUGLAS, DJ
通讯作者：
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影响因子：
4.8
作者：
Ashcroft, AE;Brinker, A;Radford, SE
通讯作者：
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影响因子：
4.8
作者：
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通讯作者：
Ashcroft, AE