Characterization of native protein structure with ion mobility mass spectrometry, multiplexed fragmentation strategies and multivariant analysis
Characterization of native protein structure with ion mobility mass spectrometry, multiplexed fragmentation strategies and multivariant analysis
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DOI：
10.1016/j.ijms.2021.116588
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发表时间：
2021-03-26
影响因子：
1.8
通讯作者：
Barran, Perdita E.
中科院分区：
文献类型：
--
作者：
Black, Rachelle;Barkhanskiy, Alexey;Barran, Perdita E.
活化离子迁移率测量提供了对蛋白质三级和四级结构稳定性的见解，并且当与解离策略结合时，可以描绘折叠是如何被破坏的。在这项工作中，我们使用213 nm的光解离加上离子迁移质谱和碰撞活化探测模型蛋白质的构象景观。UVPD实验在源活化后的蛋白质上和在离子迁移率分离后的碰撞活化的光产物上进行。对于所有观察到的细胞色素c的构象，有一个显着增加的UVPD碎片产量与另外的碰撞活化后的流动性。多聚体蛋白质、伴刀豆球蛋白a和血红蛋白也采用了类似的策略。对于这些复合物，CID导致亚基产物中的“经典”不对称电荷分布，当在其之前进行UV照射时，从亚基内产生片段。结合这些策略提供了复杂的多维数据集，信息丰富，在这里我们用多变量分析（MVA）挖掘。这种方法很容易确定UVPD和CID片段化模式的差异作为构象的函数，并揭示了前体天然结构的诊断信息，而不受当前方法的限制，只分配骨架裂解位点。(c)2021由爱思唯尔公司出版
Activated ion mobility measurements provide insights to the stability of tertiary and quaternary structures of proteins and when coupled with dissociation strategies can delineate how the fold is disrupted. In this work, we use 213 nm photodissociation coupled with ion mobility mass spectrometry and collisional activation to probe the conformational landscape of model proteins. UVPD experiments are performed on proteins following in source activation and on collisionally activated photoproducts post ion mobility separation. For all observed conformations of cytochrome c, there is a significant increase in the UVPD fragmentation yield with the addition of collisional activation post mobility. Similar strategies are deployed with the multimeric proteins, concanavalin a, and haemoglobin. For these complexes' CID results in 'classical' asymmetric charge distribution in subunit products, which when preceded by UV irradiation, yields fragments from within sub-units. Combining these strategies provides complex multidimensional datasets, rich in information, which here we mine with multivariate analysis (MVA). This approach readily determines differences in UVPD and CID fragmentation patterns as a function of conformation and reveals diagnostic information about the precursor native structure without the limitations of current methods that only assign backbone cleavage sites.(c) 2021 Published by Elsevier B.V.