Biofabrication of functional protein nanoparticles through simple His-tag engineering.

Hèctor López-Laguna,Laura Sánchez-García,Eloi Parladé,Nerea Roher,Antonio Villaverde,Sandra Flores,Olivia Cano-Garrido,Verónica Nolan,Eric Voltà-Durán,Alejandro Sánchez-Chardi,Julieta M Sánchez,Neus Ferrer-Miralles,Mauricio Rojas-Peña,José Vicente Carratalá,Ario De Marco,Esther Vazquez,Ugutz Unzueta,Naroa Serna

doi:10.1021/acssuschemeng.1c04256

Abstract

We have developed a simple, robust, and fully transversal approach for the a-la-carte fabrication of functional multimeric nanoparticles with potential biomedical applications, validated here by a set of diverse and unrelated polypeptides. The proposed concept is based on the controlled coordination between Zn2+ ions and His residues in His-tagged proteins. This approach results in a spontaneous and reproducible protein assembly as nanoscale oligomers that keep the original functionalities of the protein building blocks. The assembly of these materials is not linked to particular polypeptide features, and it is based on an environmentally friendly and sustainable approach. The resulting nanoparticles, with dimensions ranging between 10 and 15 nm, are regular in size, are architecturally stable, are fully functional, and serve as intermediates in a more complex assembly process, resulting in the formation of microscale protein materials. Since most of the recombinant proteins produced by biochemical and biotechnological industries and intended for biomedical research are His-tagged, the green biofabrication procedure proposed here can be straightforwardly applied to a huge spectrum of protein species for their conversion into their respective nanostructured formats.

Highlights

Protein materials result from the controlled self-assembly of individual polypeptides under defined architectural patterns.[1,2] In contrast to other chemical categories of building blocks, proteins generate supramolecular materials that benefit from the structural and functional capabilities and the particular versatility shown by these biomolecules
On the bases of the presented data, we propose the use of divalent cations as a universal trigger of His-tag-based protein oligomerization into the nanoscale, through a simple procedure that can be applied on demand to any His-tagged polypeptide
Other strategies based on structural selection, ensuring periodicity between particular amino acids (Glu, Cys, Asp, and His79) or through the incorporation of artificial amino acids with high affinity for cations,[80] are addressed to facilitate the formation of oligomeric structures

Summary

Introduction

Protein materials result from the controlled self-assembly of individual polypeptides under defined architectural patterns.[1,2] In contrast to other chemical categories of building blocks, proteins generate supramolecular materials that benefit from the structural and functional capabilities and the particular versatility shown by these biomolecules. Cross-molecular interactions can be conferred by self-assembling protein domains recruited from nature; when they are fused to a protein of interest, they drive the organization of the whole fusion into regular oligomers.[12,13] diverse categories of chemical cross-linkers have been explored for the construction of protein materials.[14−16] the design of materials usable in biological interfaces should be ideally achieved through selfassembly, with a minimal extent of protein engineering and without the addition of chemical couplers that are potentially toxic This would minimize perturbations of the original protein conformation, preserve the full functionality, and avoid

Methods

Results

Conclusion