High resolution time-of-flight mass analysis of the entire range of intact singly-charged proteins.
High resolution time-of-flight mass analysis of the entire range of intact singly-charged proteins.
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DOI：
10.1021/ac202001z
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发表时间：
2011-12-15
影响因子：
7.4
通讯作者：
Reilly, Peter T. A.
中科院分区：
文献类型：
--
作者：
Lee, Jeonghoon;Chen, Huijuan;Liu, Tiancheng;Berkman, Clifford E.;Reilly, Peter T. A.
提出了对任意大小的完整单电荷蛋白进行高分辨率分析的原理证明。在常压下，通过电喷雾电离和表面诱导电荷还原产生单电荷蛋白离子。入口和捕获系统“停止”蛋白质离子在一个非常宽的范围内向前动量，由大半径线性离子阱的数字产生的电场捕获，然后它们被移动到一个较小半径的线性离子阱中，并使用数字波形操作在其出口端盖电极前收集和集中。然后，根据需要，蛋白质离子在紧准直的离子束中从小半径线性四极杆的末端喷出，具有仪器定义的动能，进入正交加速反射飞行时间质量分析仪的加速区域，在那里，它们的飞行时间被Photonis双极TOF探测器测量和检测。我们提出的结果清楚地证明，大质量的单电荷离子可以产生高分辨率的质谱，具有非常低的化学噪声，并且随着整个谱的质量增加而不会失去灵敏度。链亲和素- cy5与生物素化肽模拟物结合，证明了非共价结合蛋白复合物的分析。我们的结果表明，在整个范围内的蛋白质可以直接定量使用我们的质量分析技术。我们提出的证据表明，溶剂分子非共价加合到蛋白质上，同时产生一致的飞行时间分布。最后，我们提供了对未来的展望，这将导致快速测量和量化蛋白质分布的能力。
The proof of principle for high resolution analysis of intact singly-charged proteins of any size is presented. Singly-charged protein ions were produced by electrospray ionization followed by surface-induced charge reduction at atmospheric pressure. The inlet and trapping system “stops” to forward momentum of the protein ion over a very broad range to be captured by the digitally-produced electric fields of a large radius linear ion trap whereupon they are moved into a smaller radius linear ion trap and collected and concentrated in front of its exit end cap electrode using digital waveform manipulation. The protein ions are then ejected on demand from the end of the small radius linear quadrupole in a tightly collimated ion beam with a instrumentally defined kinetic energy into the acceleration region of an orthogonal acceleration reflectron time-of-flight mass analyzer where their flight times were measured and detected with a Photonis BiPolar TOF detector. We present results that clearly prove that massive singly-charged ions can yield high resolution mass spectra with very low chemical noise and without loss of sensitivity with increasing mass across the entire spectrum. Analysis of noncovalently bound protein complexes was demonstrated with streptavidin-Cy5 bound with a biotinylated peptide mimic. Our results suggest proteins across the entire range can be directly quantified using our mass analysis technique. We present evidence that solvent molecules noncovalently adduct onto the proteins while yielding consistent flight time distributions. Finally, we provide a look into future that will result from the ability to rapidly measure and quantify protein distributions.
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影响因子：
--
作者：
Laschober, C;Kaufman, SL;Szymanski, WW
通讯作者：
Szymanski, WW
影响因子：
7.4
作者：
Chernushevich, IV;Thomson, BA
通讯作者：
Thomson, BA
影响因子：
7.4
作者：
Sobott, F;Hernández, H;Robinson, CV
通讯作者：
Robinson, CV
影响因子：
2.3
作者：
Bacher, G;Szymanski, WW;Allmaier, G
通讯作者：
Allmaier, G
影响因子：
56.9
作者：
FENN, JB;MANN, M;WHITEHOUSE, CM
通讯作者：
WHITEHOUSE, CM