Department of Chemistry, University of Wisconsin, Madison, Wisconsin 53706, United States.
J Am Soc Mass Spectrom. 2021 Jun 2;32(6):1319-1325. doi: 10.1021/jasms.0c00462. Epub 2021 Mar 23.
Top-down proteomics is a key mass spectrometry-based technology for comprehensive analysis of proteoforms. Proteoforms exhibit multiple high charge states and isotopic forms in full MS scans. The dissociation behavior of proteoforms in different charge states and subjected to different collision energies is highly variable. The current widely employed data-dependent acquisition (DDA) method selects a narrow / range (corresponding to a single proteoform charge state) for dissociation from the most abundant precursors. We describe here Mesh, a novel dissociation strategy, to dissociate multiple charge states of one proteoform with multiple collision energies. We show that the Mesh strategy has the potential to generate fragment ions with improved sequence coverage and improve identification ratios in top-down proteomic analyses of complex samples. The strategy is implemented within an open-source instrument control software program named MetaDrive to perform real time deconvolution and precursor selection.
自上而下的蛋白质组学是一种基于质谱的关键技术，可用于全面分析蛋白质形式。蛋白质形式在全 MS 扫描中表现出多种高电荷状态和同位素形式。不同电荷状态下的蛋白质形式以及在不同碰撞能量下的解离行为具有高度的可变性。目前广泛使用的数据依赖采集（DDA）方法从最丰富的前体中选择一个窄的范围（对应于单个蛋白质形式的电荷状态）进行解离。我们在这里描述了 Mesh，一种新的解离策略，用于用多种碰撞能量解离一个蛋白质形式的多个电荷状态。我们表明，Mesh 策略有可能在复杂样品的自上而下蛋白质组学分析中产生具有改进的序列覆盖率的片段离子，并提高鉴定率。该策略在名为 MetaDrive 的开源仪器控制软件程序中实现，以执行实时解卷积和前体选择。
