Uncoiling collagen: a multidimensional mass spectrometry study
Uncoiling collagen: a multidimensional mass spectrometry study
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DOI：
10.1039/c5an01757b
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发表时间：
2016-01-01
期刊：
影响因子：
4.2
通讯作者：
O'Connor, P. B.
中科院分区：
文献类型：
--
作者：
Simon, H. J.;van Agthoven, M. A.;O'Connor, P. B.
质谱法可用于通过激活前体并分析所产生的一系列碎片来确定离子的结构信息。二维傅里叶变换离子回旋共振质谱（2D FT-ICR MS）是一种在单一光谱中关联碎片和前体离子的质荷比（m/z）的技术。2D FT-ICR MS记录样品中所有离子的碎片化，而无需分离。为了分析特定的前体，采取了光谱的水平横截面（碎片离子扫描），为传统的串联质谱（MS/MS）实验提供了一种替代方法。在这项工作中，2D FT-ICR MS已被用于研究胰蛋白酶消化的I型胶原蛋白，一个大的蛋白质。从2D FT-ICR MS光谱中提取前体m/z比的碎片离子扫描：951.81、850.41、634.34和659.34，并将2D FT-ICR MS光谱与使用不同碎片化方法的一组1D MS/MS光谱进行比较。结果表明，二维质谱法在复杂混合物的MS/MS中表现出色，通过消除污染峰简化了谱图，并有助于鉴定样品中的物种。目前，使用台式计算机，2D FT-ICR MS受到数据处理能力的限制，这一限制应该使用集群并行计算来缓解。为了探索胶原蛋白的2D FT-ICR MS，在合理的计算时间内，碎片离子维度的分辨率限于256 k数据点（与1D MS/MS光谱中的4 M数据点相比），但垂直前体离子维度有4096条线，因此总数据集为1G数据点（4Gbytes）。用盲法、未优化的2D FT-ICR MS实验获得的碎片离子覆盖率低于常规MS/MS，但无论选择和分离如何，都可获得样品中所有离子的MS/MS信息。最后，虽然所有2D FT-ICR MS峰归属都是在1D FT-ICR MS数据的帮助下进行的，但这些结果证明了2D FT-ICR MS作为研究复杂蛋白质消化混合物的技术的前景。
Mass spectrometry can be used to determine structural information about ions by activating precursors and analysing the resulting series of fragments. Two-dimensional Fourier transform ion cyclotron resonance mass spectrometry (2D FT-ICR MS) is a technique that correlates the mass-to-charge (m/z) ratio of fragment and precursor ions in a single spectrum. 2D FT-ICR MS records the fragmentation of all ions in a sample without the need for isolation. To analyse specific precursors, horizontal cross-sections of the spectrum (fragment ion scans) are taken, providing an alternative to conventional tandem mass spectrometry (MS/MS) experiments. In this work, 2D FT-ICR MS has been used to study the tryptic digest of type I collagen, a large protein. Fragment ion scans have been extracted from the 2D FT-ICR MS spectrum for precursor m/z ratios: 951.81, 850.41, 634.34, and 659.34, and 2D FT-ICR MS spectra are compared with a set of 1D MS/MS spectra using different fragmentation methods. The results show that two-dimensional mass spectrometry excells at MS/MS of complex mixtures, simplifying spectra by eliminating contaminant peaks, and aiding the identification of species in the sample. Currently, with desktop computers, 2D FT-ICR MS is limited by data processing power, a limitation which should be alleviated using cluster parallel computing. In order to explore 2D FT-ICR MS for collagen, with reasonable computing time, the resolution in the fragment ion dimension is limited to 256k data points (compared to 4M data points in 1D MS/MS spectra), but the vertical precursor ion dimension has 4096 lines, so the total data set is 1G data points (4 Gbytes). The fragment ion coverage obtained with a blind, unoptimized 2D FT-ICR MS experiment was lower than conventional MS/MS, but MS/MS information is obtained for all ions in the sample regardless of selection and isolation. Finally, although all 2D FT-ICR MS peak assignments were made with the aid of 1D FT-ICR MS data, these results demonstrate the promise of 2D FT-ICR MS as a technique for studying complex protein digest mixtures.