Design, synthesis, and characterization of protein origami based on self-assembly of a brick and staple artificial protein pair.

Fiche publication

Date publication

mars 2023

Journal

Proceedings of the National Academy of Sciences of the United States of America

Auteurs

Membres identifiés du Cancéropôle Est :
Dr DUJARDIN Erik

Tous les auteurs :
Moreaud L, Viollet S, Urvoas A, Valerio-Lepiniec M, Mesneau A, Li de la Sierra-Gallay I, Miller J, Ouldali M, Marcelot C, Balor S, Soldan V, Meriadec C, Artzner F, Dujardin E, Minard P

Lien Pubmed

https://www.ncbi.nlm.nih.gov/pubmed/36893280

Résumé

A versatile strategy to create an inducible protein assembly with predefined geometry is demonstrated. The assembly is triggered by a binding protein that staples two identical protein bricks together in a predictable spatial conformation. The brick and staple proteins are designed for mutual directional affinity and engineered by directed evolution from a synthetic modular repeat protein library. As a proof of concept, this article reports on the spontaneous, extremely fast and quantitative self-assembly of two designed alpha-repeat (αRep) brick and staple proteins into macroscopic tubular superhelices at room temperature. Small-angle X-ray scattering (SAXS) and transmission electron microscopy (TEM with staining agent and cryoTEM) elucidate the resulting superhelical arrangement that precisely matches the a priori intended 3D assembly. The highly ordered, macroscopic biomolecular construction sustains temperatures as high as 75 °C thanks to the robust αRep building blocks. Since the α-helices of the brick and staple proteins are highly programmable, their design allows encoding the geometry and chemical surfaces of the final supramolecular protein architecture. This work opens routes toward the design and fabrication of multiscale protein origami with arbitrarily programmed shapes and chemical functions.