De Novo Designed Peptide and Protein Hairpins Self‐Assemble into Sheets and Nanoparticles

Abstract

The design and assembly of peptide‐based materials has advanced considerably, leading to a variety of fibrous, sheet, and nanoparticle structures. A remaining challenge is to account for and control different possible supramolecular outcomes accessible to the same or similar peptide building blocks. Here a de novo peptide system is presented that forms nanoparticles or sheets depending on the strategic placement of a “disulfide pin” between two elements of secondary structure that drive self‐assembly. Specifically, homodimerizing and homotrimerizing de novo coiled‐coil α‐helices are joined with a flexible linker to generate a series of linear peptides. The helices are pinned back‐to‐back, constraining them as hairpins by a disulfide bond placed either proximal or distal to the linker. Computational modeling indicates, and advanced microscopy shows, that the proximally pinned hairpins self‐assemble into nanoparticles, whereas the distally pinned constructs form sheets. These peptides can be made synthetically or recombinantly to allow both chemical modifications and the introduction of whole protein cargoes as required.

Bottom-up supramolecular assembly in two dimensions

“…hydrophobic effects) are valuable orthogonal motifs that require the assistance of directional interactions to facilitate supramolecular polymerisation in 2D. Pre-folded scaffolds, such as a-helical peptide backbones 63,64 or self-assembled nanotubes, 65 can also direct monomer elongation by segregating supramolecular motifs into domains with dened geometries (e.g. Leu zippers (Fig.…”

“…For example, helical peptide and protein scaffolds have been assembled in 2D by controlling binding domains such as metal coordination groups ( Fig. 4B ), 101 oppositely charged amphiphiles, 102 hydrophobic zipping of coiled-coils 64 and trigonal orientation of orthogonal domains. 63 Protein interfaces have also produced porous 2D assemblies by geometrically restricted oligomerisation of covalent protein dimers 103 and hydrophobic interfaces.…”

Advances in Rational Protein Engineering toward Functional Architectures and Their Applications in Food Science

“…On the basis of the established principles and methods, numerous architectures ranging from nanocage particles to high-dimensional structures have been created. 43,44 Except for the construction of novel structures, custom functional structures could also be produced through de novo design. For example, by using the scaffold of the de novo protein GRα 3 D, a copper-binding motif has been introduced in the scaffold to endow the complex structure with superoxide dismutase activity.…”

“…As time went on, de novo protein design has undergone rapid development due to powerful computational capabilities and advanced algorithms, and a reliable design routine has been established for accurate design of a protein structure. , The process of de novo design mainly involves the following steps: (1) definition of the target architecture or topology, (2) generation of suitable protein backbones and sampling of side-chain conformations at low energy, and (3) a compatibility evaluation of the designed sequence and structure. On the basis of the established principles and methods, numerous architectures ranging from nanocage particles to high-dimensional structures have been created. , Except for the construction of novel structures, custom functional structures could also be produced through de novo design. For example, by using the scaffold of the de novo protein GRα 3 D, a copper-binding motif has been introduced in the scaffold to endow the complex structure with superoxide dismutase activity .…”

Tubular Assembly Formation Induced by Leucine Alignment along the Hydrophobic Helix of Amphiphilic Polypeptides

“…Introducing a well-designed secondary structure with a specific interaction into an amphipathic molecule results in the design of an artificial peptide assembly that is precisely morphologically controlled. Woolfson and coworkers succeeded in designing a spherical structure in which helices were regularly arranged using a coiled-coil formation, which is a typical interaction between α-helices [ 2 , 3 , 4 ]. Conticello and coworkers prepared artificial nanotubes with uniform diameters by rational design of the orientation, inclination, and packing of the α-helices using the spatial arrangement of amino acids on the helix [ 5 , 6 ].…”