Val-NCA-mediated
acylation of amino acids and peptides. (a) Schematic
of Val-NCA-mediated peptide acylation in the presence of either oleic
acid or octanoic acid. (b–e) Overlay of extracted high-resolution
mass spectrometry (HRMS) spectra ([M – H]−1) for peptide or acyl peptide products observed in 24 h incubated
reaction mixtures starting from Val-NCA (20 mM), tryptophan (W) (20
mM), EPPS (300 mM), and either oleic acid (20 mM) (b and c) or octanoic
acid (20 mM) (d and e). For similar HRMS analyses of the oligo-valine
products (Vn) [R2 = CH(CH3)2], see Figure S1, SI). HRMS experiments were carried out in negative
mode. Panels (b) and (d) highlight the W-containing native peptides
observed in the presence of oleic acid and octanoic acid, respectively.
Panels (c) and (e) highlight the W-containing oleoylated (Ol-) and
octanoylated (Oc-) products, respectively. See the SI for tabulation of mass errors and an HPLC-extracted ion
chromatogram showing the product distribution observed in the case
of oleic acid (b, c).