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用于蛋白质组学研究的双线性离子阱-傅里叶变换离子回旋共振质谱仪的性能评估。

Performance evaluation of a dual linear ion trap-Fourier transform ion cyclotron resonance mass spectrometer for proteomics research.

机构信息

Department of Genome Sciences, University of Washington, Seattle, WA, USA.

出版信息

J Proteomics. 2013 Aug 2;88:109-19. doi: 10.1016/j.jprot.2013.04.009. Epub 2013 Apr 13.

Abstract

UNLABELLED

A novel dual cell linear ion trap Fourier transform ion cyclotron resonance mass spectrometer (FT-ICR MS) and its performance characteristics are reported. A linear ion trap-Fourier transform ion cyclotron resonance mass spectrometer has been modified to incorporate a LTQ-Velos mass spectrometer. This modified instrument features efficient ion accumulation and fast MS/MS acquisition capabilities of dual cell linear RF ion trap instruments coupled to the high mass accuracy, resolution, and dynamic range of a FT-ICR for improved proteomic coverage. The ion accumulation efficiency is demonstrated to be an order of magnitude greater than that observed with LTQ-FT Ultra instrumentation. The proteome coverage with yeast was shown to increase over the previous instrument generation by 50% (100% increase on the peptide level). In addition, many lower abundance level yeast proteins were only detected with this modified instrument. This novel configuration also enables beam type CID fragmentation using a dual cell RF ion trap mass spectrometer. This technique involves accelerating ions between traps while applying an elevated DC offset to one of the traps to accelerate ions and induce fragmentation. This instrument design may serve as a useful option for labs currently considering purchasing new instrumentation or upgrading existing instruments.

BIOLOGICAL SIGNIFICANCE

A novel hybrid mass spectrometer that allows increased MS/MS acquisition rates with high mass measurement accuracy and new ion fragmentation methods greatly improves the number of proteins, posttranslational modifications and protein-protein interactions that can be identified from cells.

摘要

未标记

本文报道了一种新型双池线性离子阱傅里叶变换离子回旋共振质谱仪(FT-ICR MS)及其性能特点。对线性离子阱-傅里叶变换离子回旋共振质谱仪进行了改装,使其与 LTQ-Velos 质谱仪相结合。该改装仪器具有高效的离子积累和快速的 MS/MS 采集能力,适用于双池线性 RF 离子阱仪器,结合了 FT-ICR 的高质量精度、分辨率和动态范围,可提高蛋白质组覆盖范围。离子积累效率比 LTQ-FT Ultra 仪器观察到的效率高一个数量级。与前一代仪器相比,酵母的蛋白质组覆盖范围增加了 50%(肽水平增加 100%)。此外,许多低丰度水平的酵母蛋白仅用这种改良仪器检测到。这种新型配置还可在双池 RF 离子阱质谱仪中使用束型 CID 碎裂技术。该技术涉及在两个阱之间加速离子,同时对其中一个阱施加升高的直流偏置,以加速离子并诱导碎裂。这种仪器设计可能是当前考虑购买新仪器或升级现有仪器的实验室的一个有用选择。

生物学意义

这种新型混合质谱仪能够以高质量测量精度和新的离子碎裂方法提高 MS/MS 采集速率,大大增加了可从细胞中鉴定的蛋白质、翻译后修饰和蛋白质-蛋白质相互作用的数量。

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