Large scale localization of protein phosphorylation by use of electron capture dissociation mass spectrometry.
We used on-line electron capture dissociation (ECD) for the large scale identification and localization of sites of phosphorylation. Each FT-ICR ECD event was paired with a linear ion trap collision-induced dissociation (CID) event, allowing a direct comparison of the relative merits of ECD and CID for phosphopeptide identification and site localization. Linear ion trap CID was shown to be most efficient for phosphopeptide identification, whereas FT-ICR ECD was superior for localization of sites of phosphorylation. The combination of confident CID and ECD identification and confident CID and ECD localization is particularly valuable in cases where a phosphopeptide is identified just once within a phosphoproteomics experiment.
Sweet
Steve M M
SM
Cancer Research UK Growth Factor Group, College of Life and Environmental Sciences, Universityof Birmingham, Edgbaston, Birmingham B152TT, United Kingdom.
Bailey
Christopher M
CM
Cunningham
Debbie L
DL
Heath
John K
JK
Cooper
Helen J
HJ
D000595:Amino Acid Sequence; D000818:Animals; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D010748:Phosphopeptides / Q000032:analysis / Q000737:chemistry; D010766:Phosphorylation; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D015203:Reproducibility of Results
D010748:Phosphopeptides; D011506:Proteins