Increased chemical and topological diversity would enhance ligand engineering to elicit precise biological activity, particularly against challenging targets. We have engineered hybrids of fibronectin domains and small molecules to merge the benefits of distinct pharmacophore activities with engineerable, high-affinity proteins. The protein-small molecule (PriSM) hybrids are engineered via yeast display in which a combinatorial protein library, each with a single cysteine, is coupled to acetazolamide via a maleimide-PEG linker and sorted for strong, selective binders via flow cytometry. The resultant PriSMs demonstrate potent, isoform-specific inhibition of carbonic anhydrase with 80-fold increase in specificity and 9-fold potency gain relative to the protein or pharmacophore components alone. The platform flexibility is demonstrated via engineering PriSMs with variable conjugation sites, linker lengths, and isoform specificities. Enhanced molecular diversity via engineered PriSMs advance discovery of potent, specific ligands.

Engineered protein-small molecule hybrids empower selective inhibition

ACS Spring 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.