Improved Protein and PTM Characterization with a Practical Electron-Based Fragmentation on Q-TOF Instruments.
Improved Protein and PTM Characterization with a Practical Electron-Based Fragmentation on Q-TOF Instruments.
复制标题
改善了基于Q-TOF仪器的实用电子碎片，改善了蛋白质和PTM表征。
DOI：
10.1021/jasms.0c00482
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发表时间：
2021-08-04
影响因子：
3.2
通讯作者：
Stafford G
中科院分区：
文献类型：
--
作者：
Beckman JS;Voinov VG;Hare M;Sturgeon D;Vasil'ev Y;Oppenheimer D;Shaw JB;Wu S;Glaskin R;Klein C;Schwarzer C;Stafford G
基于电子的解离 (ExD) 产生完整蛋白质的整洁质谱图，同时保留不稳定的翻译后修饰。然而，技术挑战将该选项限制为仅少数高端质谱仪。我们开发了一种高效的 ExD 池，可以在不到一小时的时间内改装到当前的 LC/Q-TOF 仪器中。已开发支持软件来采集、处理和注释肽和蛋白质 ExD 碎片光谱。除了产生互补断裂之外，ExD 光谱还能够通过侧链断裂来区分许多同量异位亮氨酸/异亮氨酸和异天冬氨酸/天冬氨酸对。 ExD 细胞保留磷酸化和糖基化修饰。它还可以更有效地片段化较长的肽，以揭示同一蛋白链上多个翻译后修饰之间的信号串扰，并裂解胱氨酸打结蛋白和完整抗体中的二硫键。 ExD 细胞将碰撞激活与电子断裂相结合的能力，可以通过破坏分子内静电相互作用来实现更完整的序列覆盖，而分子内静电相互作用可以将大肽和蛋白质的片段固定在一起。 ExD 细胞实现的这些增强功能扩大了可分析的肽和蛋白质的大小，以及表征其翻译后修饰的分析确定性。
Electron-based dissociation (ExD) produces uncluttered mass spectra of intact proteins while preserving labile post-translational modifications. However, technical challenges have limited this option to only a few high-end mass spectrometers. We have developed an efficient ExD cell that can be retrofitted in less than an hour into current LC/Q-TOF instruments. Supporting software has been developed to acquire, process, and annotate peptide and protein ExD fragmentation spectra. In addition to producing complementary fragmentation, ExD spectra enable many isobaric leucine/isoleucine and isoaspartate/aspartate pairs to be distinguished by side-chain fragmentation. The ExD cell preserves phosphorylation and glycosylation modifications. It also fragments longer peptides more efficiently to reveal signaling cross-talk between multiple post-translational modifications on the same protein chain and cleaves disulfide bonds in cystine knotted proteins and intact antibodies. The ability of the ExD cell to combine collisional activation with electron fragmentation enables more complete sequence coverage by disrupting intramolecular electrostatic interactions that can hold fragments of large peptides and proteins together. These enhanced capabilities made possible by the ExD cell expand the size of peptides and proteins that can be analyzed as well as the analytical certainty of characterizing their post-translational modifications.
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影响因子：
4.4
作者：
Hoopmann MR;Kusebauch U;Palmblad M;Bandeira N;Shteynberg DD;He L;Xia B;Stoychev SH;Omenn GS;Weintraub ST;Moritz RL
通讯作者：
Moritz RL
影响因子：
7.4
作者：
Shaw, Jared B.;Malhan, Neha;Voinov, Valery G.
通讯作者：
Voinov, Valery G.
DOI：
10.1021/jasms.0c00484
发表时间：
2021-06-02
影响因子：
3.2
作者：
Shen X;Xu T;Hakkila B;Hare M;Wang Q;Wang Q;Beckman JS;Sun L
通讯作者：
Sun L
影响因子：
7.4
作者：
Brodbelt JS
通讯作者：
Brodbelt JS
影响因子：
7.4
作者：
Frese, Christian K.;Altelaar, A. F. Maarten;Mohammed, Shabaz
通讯作者：
Mohammed, Shabaz