Formation of phosphopeptide metal ion complexes in liquid chromatography electrospray mass spectrometry and their influence on phosphopeptide detection英文文献.pdfVIP

RAPID COMMUNICATIONS IN MASS SPECTROMETRY Rapid Commun. Mass Spectrom. 2005; 19: 2747–2756 Published online in Wiley InterScience (). DOI: 10.1002/rcm.2105 Formation of phosphopeptide-metal ion complexes in liquid chromatography/electrospray mass spectrometry and their influence on phosphopeptide detection 1{ 1{ 1 2 1,2 Suya Liu , Cunjie Zhang , J. Larry Campbell , Haixia Zhang , Ken K.-C. Yeung , Victor K. M. Han1 and Gilles A. Lajoie1,2* 1Department of Biochemistry, University of Western Ontario, London, Ontario, N6A 5B7, Canada 2Department of Chemistry, University of Western Ontario, London, Ontario, N6A 5B7, Canada Received 30 May 2005; Revised 19 July 2005; Accepted 19 July 2005 Despite major advances in mass spectrometry, the detection of phosphopeptides by liquid chroma- tography with electrospray mass spectrometry (LC/ES-MS) still remains very challenging in proteo- mics analysis. Phosphopeptides do not protonate efficiently due to the presence of one or more acidic phosphate groups, making their detection difficult. However, other mechanisms also contri- bute to the difficulties in phosphopeptide analysis by LC/ES-MS. We report here on one such undo- cumented problem: the formation of phosphopeptide-metal ion complexes during LC/ES-MS. It is demonstrated that both synthetic phosphopeptides and phosphopeptides from bovine b-casein and a-casein form phosphopeptide-metal ion complexes containing iron and aluminum ions, resulting in a dramatic decrease in signal intensity of the protonated phosphopeptides. The interaction of phosphopeptides with metal ions on the surface of the C18 stationary phase is also shown to alter their chromatographic behavior o