A novel precursor ion discovery method on a hybrid quadrupole orthogonal acceleration time-of-flight (Q-TOF) mass spectrometer for studying protein phosphorylation
A novel precursor ion discovery method on a hybrid quadrupole orthogonal acceleration time-of-flight (Q-TOF) mass spectrometer for studying protein phosphorylation
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DOI：
10.1016/s1044-0305(02)00420-8
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发表时间：
2002-07-01
影响因子：
3.2
通讯作者：
Vissers, JPC
中科院分区：
文献类型：
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作者：
Bateman, RH;Carruthers, R;Vissers, JPC
串联四极杆飞行时间（Q-TOF）质谱仪已被编程，使磷酸化的肽可以自动发现和识别的方式类似于使用前体离子或中性损失扫描，但不需要扫描的四极杆质量过滤器。相反，该方法利用Q-TOF的固有能力来快速记录质谱和产物离子谱，具有良好的灵敏度和良好的质量准确度。交替质谱，有和没有碎片，记录在高和低碰撞能量与四极杆在宽带模式下操作。该分析方法与液相色谱法兼容并依赖于液相色谱法来分离复杂混合物。该方法已通过寻找磷酸丝氨酸和磷酸苏氨酸残基中98 Da（H（3）PO（4））的中性丢失或磷酸酪氨酸中m/z 216处的磷酸化亚铵离子得到证实。该方法还包括从任何候选前体离子获取产物离子光谱，从而允许确认中性损失或产物离子，并提供额外的序列信息以帮助鉴定蛋白质并分配磷酸化位点。
A tandem quadrupole time-of-flight (Q-TOF) mass spectrometer has been programmed such that phosphorylated peptides can automatically be discovered and identified in a way similar to that of the use of precursor ion or neutral loss scanning, but without the need to scan the quadrupole mass filter. Instead, the method capitalizes on the innate capability of the Q-TOF to record mass spectra and product ion spectra quickly, with good sensitivity and with good mass accuracy. Alternate mass spectra, with and without fragmentation, are recorded at high and low collision energy with the quadrupole operating in wideband mode. The method of analysis is both compatible with and dependant on liquid chromatography for separation of complex mixtures. The method has been demonstrated by searching for the neutral loss of 98 Da (H(3)PO(4)) from phosphoserine and phosphothreonine residues, or for the phosphorylated immonium ion at m/z 216 from phosphotyrosine. The method also incorporates acquisition of the product ion spectrum from any candidate precursor ions, thereby allowing confirmation of the neutral loss or product ion and providing additional sequence information to assist identification of the protein and assign the site of phosphorylation.