Development of an Enzyme‐Inhibitor Reaction Using Cellular Retinoic Acid Binding Protein II for One‐Pot Megamolecule Assembly.

This paper presents an enzyme building block for the assembly of megamolecules. The system is based on the inhibition of the human‐derived cellular retinoic acid binding protein II (CRABP2) domain. We synthesized a synthetic retinoid bearing an arylfluorosulfate group, which uses sulfur fluoride exchange click chemistry to covalently inhibit CRABP2. We conjugated both the inhibitor and a fluorescein tag to an oligo(ethylene glycol) backbone and measured a second‐order rate constant for the protein inhibition reaction of approximately 3,600 M−1s−1. We used this new enzyme‐inhibitor pair to assemble multi‐protein structures in one‐pot reactions using three orthogonal assembly chemistries to demonstrate exact control over the placement of protein domains within a single, homogeneous molecule. This work enables a new dimension of control over specificity, orientation, and stoichiometry of protein domains within atomically precise nanostructures. [ABSTRACT FROM AUTHOR]