Desalted peptides were resuspended in 0.1% (v/v) formic acid and loaded onto HPLC-MS/MS system for analysis on an Orbitrap Q-Exactive Exploris 480 (Thermo Fisher Scientific) mass spectrometer coupled to an Easy nanoLC 1000 (Thermo Fisher Scientific) with a flow rate of 250 nl/min. The stationary phase buffer was 0.5 % formic acid and mobile phase buffer was 0.5 % (v/v) formic acid in acetonitrile. Chromatography for peptide separation was performed using increasing organic proportion of acetonitrile (5 – 40 % (v/v)) over a 120 min gradient) on a self-packed analytical column using PicoTip™ emitter (New Objective, Woburn, MA) using Reprosil Gold 120 C-18, 1.9 μm particle size resin (Dr. Maisch, Ammerbuch-Entringen, Germany). The mass spectrometry analyzer operated in data dependent acquisition mode with a top ten method at a mass range of 300–2000 Da. Data were processed using MaxQuant software (version 1.5.2.8) 58 (link) with the following setting: oxidized methionine residues and protein N-terminal acetylation as variable modification, cysteine carbamidomethylation as fixed modification, first search peptide tolerance 20 ppm, main search peptide tolerance 4.5 ppm. Protease specificity was set to trypsin with up to 2 missed cleavages allowed. Only peptides longer than five amino acids were analyzed, and the minimal ratio count to quantify a protein is 2 (proteome only). The false discovery rate (FDR) was set to 1% for peptide and protein identifications. Database searches were performed using the Andromeda search engine integrated into the MaxQuant environment 59 (link) against the UniProt Trypanosoma cruzi strain CL Brener (352153) database containing 19,242 entries (March 2020). “Matching between runs” algorithm with a time window of 0.7 min was employed to transfer identifications between samples processed using the same nanospray conditions. Protein tables were filtered to eliminate identifications from the reverse database and common contaminants.