Fig. 2

Schematic workflow used to investigate the B. cereus thiol proteome. 1/Samples were harvested from the bioreactor at early- (EEP), mid- (MEP) and late-exponential growth phase (LEP); 2/Proteins were extracted and thiol-disulfide exchanges were quenched by addition of TCA; 3/Denatured proteins were obtained; 4/Proteins with reactive Cys residues were labeled using two strategies. IDN) Reduced-Cys thiol (SH) were blocked with IAM; oxidized Cys thiols (S-OX) were then reduced with DTT and alkylated with NEM. IN) Reduced-Cys thiol were alkylated with IAM and remaining non-alkylated Cys thiols were blocked with NEM; 5/Proteins were digested with trypsin; 6/Peptides were analyzed by LC-MS/MS; 7/Modified Cys were identified; 8/ Proteins were identified and the thiol proteome profiled. TCA: trichloroacetic acid; IAM: iodoacetamide; DTT: dithiothreitol; NEM: N-ethylmaleimide