Improved Peak Detection and Deconvolution of Native Electrospray Mass Spectra from Large Protein Complexes.
Improved Peak Detection and Deconvolution of Native Electrospray Mass Spectra from Large Protein Complexes.
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DOI：
10.1007/s13361-015-1235-6
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发表时间：
2015-12
影响因子：
3.2
通讯作者：
Guan, Shenheng
中科院分区：
文献类型：
--
作者：
Lu, Jonathan;Trnka, Michael J.;Roh, Soung-Hun;Robinson, Philip J. J.;Shiau, Carrie;Fujimori, Danica Galonic;Chiu, Wah;Burlingame, Alma L.;Guan, Shenheng
天然电喷雾电离质谱（天然MS）在保留蛋白质三级和四级结构的大部分方面的条件下测量生物分子，从而能够直接表征大型完整蛋白质组装体。然而，来自这些组件的天然光谱通常部分地被低信噪比以及由于电喷雾过程后的残余溶剂化和加合而导致的宽峰形状所掩盖。宽的峰宽以及来自高电荷离子的连续电荷状态系列是紧密间隔的事实意味着包含多个物种的原生光谱通常遭受高度的峰重叠或者包含高度交错的电荷包络。这种情况对峰检测、正确的电荷状态和电荷包络分配以及最终提取所研究的非共价组装物的相关基础质量值提出了挑战。在这份报告中，我们描述了一个全面的算法开发解决峰检测，峰重叠，并在本地质谱，称为PeakSeeker的电荷状态分配。通过检查原始质谱的二阶导数检测重叠峰。通过将电荷包络的线性组合拟合到整体实验质谱来确定分子种类的电荷状态分布。该当前软件能够对大蛋白组装体的异质、复杂和有噪声的天然质谱进行去卷积，如通过分析（a）含有严重重叠峰的合成单核体、（B）具有许多紧密交错的离子信号的RNA聚合酶II/a-鹅膏蕈碱复合物和（c）含有高水平背景噪声的人TriC复合物所证明的。
Native electrospray-ionization mass spectrometry (native MS) measures biomolecules under conditions that preserve most aspects of protein tertiary and quaternary structure, enabling direct characterization of large intact protein assemblies. However, native spectra derived from these assemblies are often partially obscured by low signal-to-noise as well as broad peak shapes due to residual solvation and adduction after the electrospray process. The wide peak widths together with the fact that sequential charge state series from highly charged ions are closely spaced means that native spectra containing multiple species often suffer from high degrees of peak overlap or else contain highly interleaved charge envelopes. This situation presents a challenge for peak detection, correct charge state and charge envelope assignment, and ultimately extracting the relevant underlying mass values of the non-covalent assemblages being investigated. In this report we describe a comprehensive algorithm developed for addressing peak detection, peak overlap, and charge state assignment in native mass spectra, called PeakSeeker. Overlapped Peaks are detected by examination of the second derivative of the raw mass spectrum. Charge state distributions of the molecular species are determined by fitting linear combinations of charge envelopes to the overall experimental mass spectrum. This current software is capable of deconvoluting heterogeneous, complex, and noisy native mass spectra of large protein assemblies as demonstrated by analysis of (a) synthetic mononucleosomes containing severely overlapping peaks, (b) an RNA polymerase II/a-amanitin complex with many closely interleaved ion signals, and (c) human TriC complex containing high levels of background noise.
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影响因子：
5.6
作者：
Lowary, PT;Widom, J
通讯作者：
Widom, J
DOI：
10.1073/pnas.37.4.205
发表时间：
1951-01-01
影响因子：
11.1
作者：
PAULING, L;COREY, RB;BRANSON, HR
通讯作者：
BRANSON, HR
影响因子：
48
作者：
Rose, Rebecca J.;Damoc, Eugen;Heck, Albert J. R.
通讯作者：
Heck, Albert J. R.
影响因子：
1.9
作者：
BONOLI, C
通讯作者：
BONOLI, C
影响因子：
--
作者：
Shiau, Carrie;Trnka, Michael J.;Fujimori, Danica Galonic
通讯作者：
Fujimori, Danica Galonic