CoA-SH is a maleimide-reactive building block which can be used for the one-step synthesis of CoA substrates from maleimide-containing precursors. This building block allows you to make custom CoA substrates for labeling ACP-tag or MCP-tag fusion proteins for a wide range of applications.
A typical reaction is performed at 30°C overnight by using N,N-dimethylformamide (DMF) and Tris-buffer (pH 7.5) as solvents. Equimolar quantities of CoA-SH and reactive maleimide are recommended.
Example reaction: Coupling of CoA-SH to a Maleimide-reactive Precursor
Figure 2. Coupling of CoA-SH to a maleimide-reactive precursor. A solution of a maleimide derivative (1 eq, 6.6 μmol) in DMF (450 μl) was added to a solution of CoA-SH (6.2 mg, 1.2 eq, 7.9 μmol) in Tris-buffer (pH 7.5, 50 μl). The reaction mixture was shaken overnight at 30°C. The solvent was evaporated under vacuum. The solid was dissolved in 1 ml water/acetonitrile (9:1, 0.08% TFA) and purified by HPLC. HPLC fractions containing the product were neutralized with NH4OH (30% aquaeous solution) and lyophilized.