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Application of Tandem Two-Dimensional Mass Spectrometry for Top-Down Deep Sequencing of Calmodulin
Journal of the American Society for Mass Spectrometry ( IF 3.1 ) Pub Date : 2018-06-04 , DOI: 10.1007/s13361-018-1978-y
Federico Floris 1 , Lionel Chiron 2 , Alice M. Lynch 1 , Mark P. Barrow 1 , Marc-André Delsuc 2, 3 , Peter B. O’Connor 1
Affiliation  

Two-dimensional mass spectrometry (2DMS) involves simultaneous acquisition of the fragmentation patterns of all the analytes in a mixture by correlating their precursor and fragment ions by modulating precursor ions systematically through a fragmentation zone. Tandem two-dimensional mass spectrometry (MS/2DMS) unites the ultra-high accuracy of Fourier transform ion cyclotron resonance (FT-ICR) MS/MS and the simultaneous data-independent fragmentation of 2DMS to achieve extensive inter-residue fragmentation of entire proteins. 2DMS was recently developed for top-down proteomics (TDP), and applied to the analysis of calmodulin (CaM), reporting a cleavage coverage of about ~23% using infrared multiphoton dissociation (IRMPD) as fragmentation technique. The goal of this work is to expand the utility of top-down protein analysis using MS/2DMS in order to extend the cleavage coverage in top-down proteomics further into the interior regions of the protein. In this case, using MS/2DMS, the cleavage coverage of CaM increased from ~23% to ~42%.

Open image in new windowGraphical Abstract
Graphical Abstract

Two-dimensional mass spectrometry, when applied to primary fragment ions from the source, allows deep-sequencing of the protein calmodulin.



中文翻译:

串联二维质谱在钙调蛋白自上而下深度测序中的应用

二维质谱(2DMS)涉及通过混合物中的所有分析物的前体离子和碎片离子的相关联,同时通过碎片区域系统地调节前体离子,从而同时获取混合物中所有分析物的碎片化模式。串联二维质谱(MS / 2DMS)将傅里叶变换离子回旋共振(FT-ICR)MS / MS的超高精度与2DMS的同时独立于数据的片段化结合在一起,从而实现整个蛋白质的广泛残基片段化。2DMS最近被开发用于自上而下的蛋白质组学(TDP),并应​​用于钙调蛋白(CaM)的分析,报告了使用红外多光子解离(IRMPD)作为裂解技术的裂解覆盖率约为23%。这项工作的目的是扩大使用MS / 2DMS进行自上而下的蛋白质分析的用途,以便将自上而下的蛋白质组学中的裂解范围进一步扩展到蛋白质的内部区域。在这种情况下,使用MS / 2DMS,CaM的裂解覆盖率从〜23%增加到〜42%。

在新窗口中打开图像图形概要
图形概要

当将二维质谱应用于源中的原始碎片离子时,可以对钙调蛋白进行深度测序。

更新日期:2018-06-04
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