5.7 Reduction, Alkylation, and Glu-C digest of V. odorata library

V. odorata peptide library fractions were reduced with 10 mM dithiothreitol (Sigma-Aldrich) at 45 °C, 850 rpm, 30 min, and alkylated with 100 mM iodoacetamide (Sigma –Aldrich) at 25 °C, 850 rpm, 15 min. Pierce C18 spin columns (Thermo Scientific) were used to remove interfering contaminants from the V. odorata samples, which were subsequently dried down in a vacuum concentrator and resuspended in acidified water (0.1% formic acid) for LC-MS/MS analysis. Digested V. odorata samples were reduced and alkylated as described above, then incubated 1:50 enzyme: substrate with endoproteinase Glu-C enzyme (Sigma) at 37 °C for 3 h.