2.5 Database search and quantitative proteomic analysis
The raw data were processed by Proteome DiscovererTM(version 2.4, Thermo Fisher, Waltham, MA, USA) and then identified by
Protein Pilot Software 5.0 (AB SCIEA, USA) in combination with SwissProt
(version 2021.06)/UniProt (version 2021.09) databases. The search
parameters are as follows: cys alkylation, iodoacetamide; static
modification, carbamidomethyl; dynamic modification, oxidation of
acetyl; proteolytic enzyme, Trypsin (Full); maximum number of missed
cleavage allowed, 2; precursor mass tolerance, 20 ppm; fragment mass
tolerance, 0.02 Da; quantitative method, TMT 6Plex. For the
quantification of TMT, the ratio of ion intensities reported by TMT in
the MS/MS spectra of the original data set was used to calculate the
fold change (FC) between samples. FC ≥ 1.2 or ≤ 0.83 and p -value
< 0.05 were set as cut-off values to designate significantly
differentially expressed proteins.