Collision-Induced Unfolding of High-m/z Native-like Protein Ions within a Trapped Ion Mobility Spectrometer

IF 2.7 2区 化学 Q2 BIOCHEMICAL RESEARCH METHODS
Olakunle O. Akinola,  and , Nicholas B. Borotto*, 
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引用次数: 0

Abstract

Native mass spectrometry (nMS) is a powerful tool for the rapid characterization of protein ions and protein–ligand complexes. By coupling nMS with ion mobility spectrometry (IMS), and collisional activation, we can rapidly obtain insights into protein conformation, and stability can be rapidly obtained. Originally incapable of this workflow, recent work enabled this collision-induced unfolding (CIU) process on commercially available Bruker timsTOF instruments. This early work, however, faced challenges in transmitting larger proteins and sought to unfold only small proteins up to 29 kDa. In this study, we continue the development of this technique and optimized instrument settings to enable the transmission of proteins up to 8,000 Th. The technique also demonstrates the capability to sufficiently energize ions to unfold native-like dimers of superoxide dismutase and β-lactoglobulin and the 45 kDa monomeric ovalbumin. When this TIMS activation technique is applied to large protein ions, however, limited unfolding was observed for bovine serum albumin, and no unfolding was observed for immunoglobulin G likely reflecting the limit of activation for this workflow.

Abstract Image

高m/z原生蛋白离子在捕获离子迁移谱仪中的碰撞诱导展开
天然质谱(nMS)是快速表征蛋白质离子和蛋白质配体复合物的有力工具。通过将nMS与离子迁移谱(IMS)和碰撞激活相结合,我们可以快速获得蛋白质构象的信息,并且可以快速获得稳定性。最初无法实现这种工作流程,但最近的工作在市售的Bruker timsTOF仪器上实现了这种碰撞诱导展开(CIU)过程。然而,这项早期工作面临着传输较大蛋白质的挑战,并试图仅展开高达29 kDa的小蛋白质。在这项研究中,我们继续发展这项技术,并优化仪器设置,使蛋白质的传输高达8000 Th。该技术还证明了充分激活离子的能力,以展开超氧化物歧化酶、β-乳球蛋白和45 kDa单体卵白蛋白的天然样二聚体。然而,当TIMS激活技术应用于大蛋白离子时,牛血清白蛋白的展开有限,而免疫球蛋白G没有展开,这可能反映了该工作流程的激活限制。
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来源期刊
CiteScore
5.50
自引率
9.40%
发文量
257
审稿时长
1 months
期刊介绍: The Journal of the American Society for Mass Spectrometry presents research papers covering all aspects of mass spectrometry, incorporating coverage of fields of scientific inquiry in which mass spectrometry can play a role. Comprehensive in scope, the journal publishes papers on both fundamentals and applications of mass spectrometry. Fundamental subjects include instrumentation principles, design, and demonstration, structures and chemical properties of gas-phase ions, studies of thermodynamic properties, ion spectroscopy, chemical kinetics, mechanisms of ionization, theories of ion fragmentation, cluster ions, and potential energy surfaces. In addition to full papers, the journal offers Communications, Application Notes, and Accounts and Perspectives
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