CRIF-ID: Development of a novel photofragmentation ion trap/time-of-flight mass spectrometer for peptide analysis

印第安纳州大学的James P. Reilly教授由化学研究仪器和设施：仪器开发（CRIF-ID）计划资助，开发用于肽测序和蛋白质鉴定的串联质谱仪。 肽片段化的选择性是通过非热活化过程实现的，该过程产生的子离子具有比通过常规方法形成的子离子更多样化的结构。 线性离子阱第一级完成离子收集和选择。 被阱选择的冷却离子在暴露于真空紫外激光时光解离。 碎片的高分辨率和精确质量分析是在反射飞行时间（TOF）第二阶段实现的。 利用该仪器，肽和蛋白质离子的真空紫外光谱和光化学倾向正在研究中，以提高测序和分析程序的速度和准确性。 肽测序和蛋白质鉴定技术对生物化学和蛋白质组学等领域至关重要。 目前的方法只能识别复杂蛋白质混合物分析中记录的相对较小部分的质谱。 他们还遇到了处理突变、翻译后修饰和基因组测序错误的困难。 正在开发的离子阱-光致碎裂-TOF仪器正在解决这些问题。 基本的进展有可能影响我们对疾病和生命过程的分子基础的理解。

Professor James P. Reilly of Indiana University is funded by the Chemistry Research Instrumentation and Facilities: Instrument Development (CRIF-ID) program to develop a tandem mass spectrometer for peptide sequencing and protein identification. Selectivity in peptide fragmentation is achieved through a non-thermal activation process which produces daughter ions that have more diverse structures than those formed by conventional approaches. A linear ion trap first stage accomplishes ion collection and selection. Cooled ions selected by the trap photodissociate upon exposure to vacuum ultraviolet laser light. High resolution and accurate mass analysis of the fragments is achieved in a reflectron time-of-flight (TOF) second stage. With this instrument the vacuum ultraviolet spectroscopy and photochemical propensities of peptide and protein ions are being investigated to improve the speed and accuracy of sequencing and analysis procedures. Peptide sequencing and protein identification technology is of central importance to fields such as biochemistry and proteomics. Current methodologies identify a relatively small fraction of the mass spectra recorded in analyses of complex protein mixtures. They also encounter difficulties coping with mutations, post-translational modifications and genome sequencing errors. The ion trap-photofragmentation-TOF instrument under development is addressing these issues. Fundamental advances have the potential to impact our understanding of the molecular basis of disease and life processes.