Helicase Activity Modulation with On-Demand Light-Based Conformational Control

Engineering a protein variant with a desired role relies on deep knowledge of the relationship between a protein's native structure and function. Using our structural understanding of a regulatory subdomain found in a family of DNA helicases, we engineered novel helicases for which the subdomain orientation is designed to switch between unwinding-inactive and -active conformations upon trans-cis isomerization of an azobenzene-based crosslinker. This on-demand light-based conformational control directly alters helicase activity as demonstrated by both bulk phase experiments and single-molecule optical tweezers analysis of one of the engineered helicases. The opto-helicase may be useful in future applications that require spatiotemporal control of DNA hybridization states.

Automated summary

Engineering a protein variant with controllable activity through light-induced conformational changes has been achieved in this study, using the structural understanding of a regulatory subdomain found in a family of DNA helicases. The designed helicase showed switchable unwinding activity upon trans-cis isomerization of an azobenzene-based crosslinker. This opto-helicase has potential applications in spatiotemporal control of DNA hybridization states.