Xuan Yue, Creese Andrew J, Horner Julie A, Cooper Helen J
Thermo Fisher Scientific, Hanna-Kunath-Str. 11, 28199 Bremen, Germany.
Rapid Commun Mass Spectrom. 2009 Jul;23(13):1963-9. doi: 10.1002/rcm.4101.
We have applied high-field asymmetric waveform ion mobility spectrometry (FAIMS) to the analysis of the phosphopeptides APLpSFRGSLPKSYVK, APLSFRGpSLPKSYVK, and APLSFRGSLPKpSYVK. The peptides have identical amino acid sequences and differ only in the site of phosphorylation. The results show that FAIMS is capable of at least partially separating these species. Separation was confirmed by coupling FAIMS with high-resolution electron transfer dissociation (ETD) mass spectrometry. Phosphorylation is retained on the ETD peptide fragments thereby allowing assignment of the site of the modification. Co-eluting phosphopeptides which differ only in the site of modification are frequently observed in liquid chromatography/tandem mass spectrometry phosphoproteomics experiments, and therefore these proof-of-principle results have implications for the application of FAIMS in that field.
我们已将高场非对称波形离子迁移谱（FAIMS）应用于磷酸化肽APLpSFRGSLPKSYVK、APLSFRGpSLPKSYVK和APLSFRGSLPKpSYVK的分析。这些肽具有相同的氨基酸序列，仅磷酸化位点不同。结果表明，FAIMS能够至少部分分离这些物种。通过将FAIMS与高分辨率电子转移解离（ETD）质谱联用，证实了分离效果。磷酸化保留在ETD肽片段上，从而能够确定修饰位点。在液相色谱/串联质谱磷酸蛋白质组学实验中，经常观察到仅修饰位点不同的共洗脱磷酸化肽，因此这些原理验证结果对FAIMS在该领域的应用具有重要意义。
