Using optimized collision energies and high resolution, high accuracy fragment ion selection to improve glycopeptide detection by precursor ion scanning
Using optimized collision energies and high resolution, high accuracy fragment ion selection to improve glycopeptide detection by precursor ion scanning
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DOI：
10.1016/s1044-0305(03)00263-0
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发表时间：
2003-07-01
影响因子：
3.2
通讯作者：
Roepstorff, P
中科院分区：
文献类型：
--
作者：
Jebanathirajah, J;Steen, H;Roepstorff, P
糖基化是最广泛的蛋白质修饰，已知可调节信号转导和几种重要的生物学相互作用。为了了解和评价糖基化的生物学作用，在特定的生物学条件下鉴定糖基化蛋白和定位糖基化位点是很重要的。为了从简单混合物中鉴定糖基化肽，即从单SIDS PAGE波段的凝胶内消化物，我们使用配备Q(2)脉冲功能的四极TOF仪器进行了高分辨率、高精度的前体离子扫描。四极杆TOF仪器的高分辨率可选择性地检测聚糖特异性片段离子，最大限度地减少了具有相同标称质量的肽衍生片段离子的干扰。前体离子扫描先前已经描述了这些聚糖衍生离子。然而，由于该方法的特异性较低，该方法的使用受到限制。使用前体离子扫描的分析可以应用于蛋白质消化的任何肽混合物，而无需事先了解蛋白质的糖基化。除了低飞摩尔（纳摩尔）检测限外，该方法的优点是不需要事先衍生化或肽混合物的酶处理。(C) 2003年美国质谱学会。
Glycosylation is the most widespread protein modification and is known to modulate signal transduction and several biologically important interactions. In order to understand and evaluate the biological role of glycosylation it is important to identify the glycosylated protein and localize the site glycosylation under particular biological conditions. To identify glycosylated peptides from simple mixtures, i.e., in-gel digests from single SIDS PAGE bands we performed high resolution, high accuracy precursor ion scanning using a quadrupole TOF instrument equipped with the Q(2) pulsing function. The high resolving power of the quadrupole TOF instrument results in the selective detection of glycan specific fragment ions minimizing the interference of peptide derived fragment ions with the same nominal mass. Precursor ion scanning has been previously described for these glycan derived ions. However the use of this method has been limited by the low specificity of the method. The analysis using precursor ion scanning can be applied to any peptide mixture from a protein digest without having previous knowledge of the glycosylation of the protein. In addition to the low femtomole (nanomolar) detection limits, this method has the advantage that no prior derivatization or enzymatic treatment of the peptide mixtures is required. (C) 2003 American Society for Mass Spectrometry.