Protein Nanotube Selectively Cleavable with DNA: Supramolecular Polymerization of “DNA-Appended Molecular Chaperones”

Abstract

Here, we report molecular chaperone GroELs that carry, at their apical domains, multiple DNA strands (ideally 28 DNA strands in total) with defined oligonucleotide (nt) sequences. This design strategy allows for the preparation of GroEL and GroEL carrying 10-nt DNA strands of 10a and 10b with complementary sequences, respectively, at their apical domains. One-dimensional coassembly of these GroELs is possible to form protein nanotube NT with an anomalous thermodynamic stability due to the exceptionally large multivalency for the coassembly. Likewise, comparably stable nanotube NT was obtained even when the apical-domain DNA strands (15c and 10d) were partially complementary to one another. Nevertheless, in sharp contrast with NT, NT, when incubated with DNA 15d, dissociates rapidly and completely because 15d preferentially hybridizes with the DNA strands of 15c in NT by displacing those of 10d, to afford a mixture of GroEL and GroEL. Even in the presence of NT, 15d cleaved off NT selectively, indicating the potential utility of NTs for targeted delivery.

Functional protein nanostructures: a chemical toolbox

“…Thereafter, a ssDNA incorporating the complementary sequence, e.g. 20-mer was used as a SL to induce supramolecular polymerization to form protein nanotubes in Tris–HCl buffer, as shown in TEM, DLS and SEC 208. The protein nanotubes were highly thermodynamically stable due to the multivalent interactions 208.…”

“…20-mer was used as a SL to induce supramolecular polymerization to form protein nanotubes in Tris–HCl buffer, as shown in TEM, DLS and SEC 208. The protein nanotubes were highly thermodynamically stable due to the multivalent interactions 208. Dissociation was achieved by adding an external ssDNA which consists of a full length complementary sequence to the 20-mer SL.…”

DNA-Functionalized Supramolecular Polymers: Dynamic Multicomponent Assemblies with Emergent Properties

“…13 Recent years have witnessed an increasing interest in hybrid molecular systems. 14,15 For example, DNA functionalization of covalent polymers provides an additional level of control on the structure and macroscopic properties of materials, allowing the construction of stimuli-responsive materials such as hydrogels and other nanomaterials, DNA-surfactants that can be applied as responsive drug delivery systems, and materials for optoelectronic devices. 16−22 Only recently have the first examples of DNA-functionalized supramolecular polymers been reported.…”

Hierarchical Assembly of DNA Origami Nanostructures Using Coiled-coil Peptides

“…coiled-coils, collagen triple helices) that impart unique functional and mechanical properties. While several examples exist of multivalent recombinant proteins covalently modified with DNA to form cages, 10 nanofibers, 11,12 and three-dimensional crystals, 13 the chemical coupling of synthetic self-assembled peptides with DNA has only recently been demonstrated. The Ke and Conticello labs demonstrated the first example of linking DNA origami nanostructures with a synthetic peptide self-assembly motif, using collagen-mimetic triple helices fused with a cationic domain to electrostatically associate with the negatively charged oligonucleotide structures.…”