Evans Caroline A, Ow Saw Yen, Smith Duncan L, Corfe Bernard M, Wright Phillip C
Department of Chemical and Biological Engineering, ChELSI Institute, University of Sheffield, Sheffield, UK.
Methods Mol Biol. 2013;981:13-23. doi: 10.1007/978-1-62703-305-3_2.
Mass spectrometry (MS)-based methods typically assess acetylation by detection of a diagnostic ion at 126.1 m/z, corresponding to the immonium ion of acetyl-lysine -NH(3), which is generated by collisionally induced dissociation. A novel implementation of this approach, based on the accurate mass and retention time technique, couples high mass resolution measurement with rapid cycling between low and elevated collision energies to generate intact and fragment high-resolution mass spectra. This allows acetyl lysine diagnostic ions at 126.1 m/z to be monitored and aligned to the precursor m/z based on retention time profile. The technique is termed Collisionally Induced Release of Acetyl Diagnostic. Sequence information is also obtained for acetylation site assignment. This technique to identify acetylation species is information independent as it does not require the sequence of the protein/peptides to identify acetylation, and thus complementary to data-dependent methods. It is suitable for analysis of acetylated peptides, or proteins enriched by immunoprecipitation with acetyl lysine-specific antibodies.
基于质谱（MS）的方法通常通过检测质荷比为126.1的诊断离子来评估乙酰化，该离子对应于乙酰赖氨酸-NH(3)的亚铵离子，它是由碰撞诱导解离产生的。这种方法的一种新应用，基于精确质量和保留时间技术，将高质量分辨率测量与低碰撞能量和高碰撞能量之间的快速循环相结合，以生成完整的和片段化的高分辨率质谱。这使得可以监测质荷比为126.1的乙酰赖氨酸诊断离子，并根据保留时间谱将其与前体质荷比进行比对。该技术被称为乙酰诊断的碰撞诱导释放。还可获得用于乙酰化位点分配的序列信息。这种识别乙酰化物种的技术是信息独立的，因为它不需要蛋白质/肽的序列来识别乙酰化，因此与数据依赖方法互补。它适用于分析乙酰化肽或用乙酰赖氨酸特异性抗体通过免疫沉淀富集的蛋白质。
