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. Author manuscript; available in PMC: 2020 Nov 20.
Published in final edited form as: Bioconjug Chem. 2019 Oct 10;30(11):2799–2804. doi: 10.1021/acs.bioconjchem.9b00550

Figure 3.

Figure 3.

Protein nucleic acid (ProNAc) conjugation catalyzed by HhC using substrate steramers. (A) Gel based validation of 1:1 ProNAc synthesis. Reactions of native-like hedgehog precursor, SHhN-DHhC, and heterologous precursor, T4Lysozyme-DHhC, at 2 μM following incubation in buffer control (NS, no sterol), and substrates: cholesterol (chol, C), II, steramers, present at initial concentration of 50 μM. Molecular weights of the conjugates (red asterisks): SHhN:S-dT10, 23.3 kDa; SHhN:S-dT20, 26.7 kDa SHhN:S-dT30, 29.4 kDa; T4Lysozyme:S-dT10, 22.8 kDa; T4Lysozyme:S-dT20, 26.1 kDa; T4Lysozyme:S-dT30, 29.4 kDa. Product DHhC is marked with a green triangle. Percent precursor consumed, %PC. (B) Conjugation kinetics from FRET assay using C−H−Y precursor. Michaelis−Menten plot (upper) and representative kinetic trace (lower). Lower plot shows reaction using 50 μM substrate and 0.1 μM C−H−Y.