Skip to main content
. 2018 Feb 23;19(4):e44981. doi: 10.15252/embr.201744981

Figure EV4. Mass spectrometry analysis of TcPINK1 autophosphorylation.

Figure EV4

  1. Intact mass spectra of TcPINK1121–570 WT or D337N expressed for different times in E. coli. The double‐edged arrows on the spectra are used to indicate a difference of 80 Da (1 phosphorylation) between peaks. “0p” indicates the theoretical mass of the unphosphorylated protein.
  2. MS/MS spectrum for the peptide 197‐212 of TcPINK1121–570 D337N, phosphorylated by GST‐TcPINK1121–570 WT (see Fig 3C). The b and y series indicate that phosphorylation takes place at Ser205. The y* series (green) show neutral loss of a phosphate group (H3PO4, −97.7 amu).
  3. Intact mass spectra of the time course phosphorylation assay of TcPINK1 D337N with CIP‐treated 15N‐TcPINK1 WT (corresponding to the plot in Fig 3D).
  4. MS/MS spectrum for the peptide 197–212 of TcPINK1121–570 14N‐D337N and 15N‐WT, after 2‐min phosphorylation (see Fig 3E). The b and y series indicate that phosphorylation takes place at Ser205 for the 14N and 15N‐labeled peptides eluting at 36.5 min, and Ser207 for the 15N‐labeled peptide eluting earlier at 35.8 min. The y* series (green) show neutral loss of a phosphate group (H3PO4, −97.7 amu). The different phosphorylation sites give rise to y(6) and y(7) ions with different masses.