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. Author manuscript; available in PMC: 2026 Jul 18.
Published in final edited form as: Nat Chem. 2025 Jul 18;17(9):1371–1382. doi: 10.1038/s41557-025-01871-3

Extended Data Figure 5. Combining HsMccB-catalyzed peptide thioester synthesis with Ubc9-catalyzed lysine acylation.

Extended Data Figure 5.

(A) Scheme for lysine acylation using an HsMccB-generated thioester and GFP with an internal minimal LACE tag sequence (IKQE). (B) Scheme for lysine acylation using an HsMccB-generated thioester and GFP with a full length LACE tag sequence (PRKVIKMESEE). (C) Optimization of peptide thioester concentration in LACE reactions. (D) Optimization of thiol concentration at pH 7.6. Excess thiol suppresses the LACE reaction. (E) Optimization of thiol concentration at pH 8.0. Excess thiol suppresses the LACE reaction, which proceeds to higher yield at pH 8.0 compared to 7.6.