Reversed-phase liquid chromatography-mass spectrometry/MS analysis
HF mass spectrometer
Briefly, a C18 EASY-spray and C18 trap columns linked to an Ultimate 3000 UPLC system (Thermo Scientific) were used for the reversed phase liquid chromatographic separations of peptides. An Q Exactive HF mass spectrometer (Thermo Scientific) was used for the subsequent mass spectra, followed by data-dependent higher-energy collisional dissociation (HCD) fragmentations from precursor ions with a charge state.