In this work, promiscuous inhibition was observed for peptides inhibiting enzymes with an observed sensitivity to enzyme and detergent concentrations. Dynamic light scattering study revealed that the peptide-enzyme aggregation occurred and was responsible for inhibiting enzyme activity. Peptide sequence analysis showed that the positively charged residues, like arginine and lysine, were crucial for inhibiting enzyme activity. To reduce peptide/enzyme aggregation, we tested phosphate-based biopolymers for recover peptide-inhibited enzyme activities. Polyphosphates and single-stranded DNA (ssDNA) were found to destabilize the aggregated complex with recovered enzyme activity. It is possible that negatively charged phosphates disrupted the electrostatic-aggregated peptide/enzyme complex. This discovery may be applied to reduce promiscuous peptide inhibition in high-throughput screening by beads library or microarrays.

Prevention of promiscuous peptides-enzyme aggregation by polyphosphate and DNA molecules

ACS Spring 2022

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