The data acquisition by LC-MS/MS was adapted from a published procedure

Derived peptides were resuspended in a loading buffer

(0.1% trifluoroacetic acid, TFA) and separated on a Water's Charged Surface

Hybrid (CSH) column (150 µm internal diameter x 15 cm; particle size: 1.7 µm).

The samples were run on an EVOSEP liquid chromatography system using the 15

samples per day preset gradient (88 min) and were monitored on a Q-Exactive

Plus Hybrid Quadrupole-Orbitrap Mass Spectrometer (Thermo Scientific).

The mass spectrometer cycle was programmed to collect one full MS scan followed by 20

data-dependent MS/MS scans. The MS scans (400-1600 m/z range, 3 x 1⁶6

AGC target, 100 ms maximum ion time) were collected at a resolution of 70,000

at m/z 200 in profile mode.

The HCD MS/MS spectra (1.6 m/z isolation width, 28%

collision energy, 1 x 1⁵5 AGC target, 100 ms maximum ion time) were

acquired at a resolution of 17,500 at m/z 200. Dynamic exclusion was set to

exclude previously sequenced precursor ions for 30 seconds. Precursor ions with

+1, and +7, +8, or higher charge states were excluded from sequencing.