采用轨道阱进行高通量中间向下分析。
High-throughput middle-down analysis using an orbitrap.
机构信息
Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20274, USA.
出版信息
J Proteome Res. 2010 Aug 6;9(8):3886-90. doi: 10.1021/pr1000994.
Abstract
This report demonstrates the application of a capillary LC-LTQ-orbitrap system to provide automated middle-down analysis of proteolytic peptides in the mass range 3000 to 10,000 Da. The novel workflow combines an underutilized method in the orbitrap-high resolution, mass-accurate product ion measurements-with software tailored to search such data (ProSightPC 2.0) and an Asp-selective chemical cleavage approach that generates peptides across an extended mass range. The strategy using high resolution mass measurements on both precursor and product ions is analogous to that widely used on FT-ICR analyzers. The approach is demonstrated in an analysis of the highly basic ribosomal proteome isolated from human MCF7 cancer cells.
摘要
本报告展示了毛细管 LC-LTQ-Orbitrap 系统在 3000 至 10000 Da 质量范围内提供自动中肽分析的应用。该新工作流程结合了一种在 Orbitrap 中未充分利用的方法——高分辨率、质量精确的产物离子测量——以及专门用于搜索此类数据的软件（ProSightPC 2.0）和一种 Asp 选择性化学裂解方法，该方法可在扩展的质量范围内生成肽。在母离子和产物离子上都使用高分辨率质量测量的策略类似于在 FT-ICR 分析仪上广泛使用的策略。该方法在分析从人 MCF7 癌细胞中分离的高度碱性核糖体蛋白质组中得到了验证。
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