Molecularly Engineered “Janus GroEL”: Application to Supramolecular Copolymerization with a Higher Level of Sequence Control

Abstract

Herein, we report the synthesis and isolation of a shape-persistent Janus protein nanoparticle derived from biomolecular machine chaperonin GroEL (A GroEL B) and its application to DNA-mediated ternary supramolecular copolymerization. For synthesizing A GroEL B with two different DNA strands A and B at its opposite apical domains, we utilized the unique biological property of GroEL, i.e., "Mg 2+ /ATPmediated ring exchange" between A GroEL A and B GroEL B with their hollow cylindrical doubledecker architectures. This exchange event was reported more than 24 years ago but has never been utilized for molecular engineering of GroEL. We leveraged DNA nanotechnology to purely isolate Janus A GroEL B and succeeded in its precision ternary supramolecular copolymerization with two DNA comonomers A** and B* that are partially complementary to A and B in A GroEL B , respectively, and programmed to self-dimerize on the other side. Transmission electron microscopy allowed us to confirm the formation of an expected dual-periodic copolymer sequence-(B*/B GroEL A/A**/A**/A GroEL B/B*)-in the form of a laterally connected lamellar assembly, rather than a single-chain copolymer.

Protein Assembly by Design

“…440 The DNA-mediated assembly strategy was also applied to generate GroEL-DNA Janus nanoparticles bearing strands with different sequences at either end of the barrel. 442 Rather than forming discrete nanotubes, these constructs polymerized into extended structures with lamellar periodic features. The GroEL barrels were presumed to adopt a hexagonal packing when combined with additional DNA strands.…”

“…In an alternative strategy for GroEL nanotube assembly, Kashiwagi et al conjugated the Cys-modified GroEL barrels to DNA strands, which enabled nanotube formation through multivalent strand complementarity . The DNA-mediated assembly strategy was also applied to generate GroEL-DNA Janus nanoparticles bearing strands with different sequences at either end of the barrel . Rather than forming discrete nanotubes, these constructs polymerized into extended structures with lamellar periodic features.…”

Intelligent Paper‐Free Sprayable Skin Mask Based on an In Situ Formed Janus Hydrogel of an Environmentally Friendly Polymer

“…It is noteworthy that Janus structures have been investigated in the fields of polymer science, soft matter and self assembly [62][63][64][65] and even some Janus hydrogels have been reported with potential to be biomedical materials etc. [66,67] Our present work distinguishes itself as the spontaneous formation of a Janus structure after spraying a solution onto skin.…”

Superstructured mesocrystals through multiple inherent molecular interactions for highly reversible sodium ion batteries

“…For example, hierarchical self-assembly via multiple noninterfering interactions on different length scales in a spatiotemporally controlled manner is the basis of various essential biological superstructures, such as supercoiled DNA ( 11 ), folded proteins ( 12 ), and biologically active cell membranes ( 13 ). Inspired by multiscale hierarchical structures in biology, substantial efforts have been devoted to developing synthesis pathways for similar intricate hierarchical systems including DNA origami construction ( 14 ), peptide-induced assembly ( 15 ), coordination-driven self-assembly ( 16 ), supramolecular polymerization ( 17 ), block copolymer assembly ( 18 ), and crystallization-driven self-assembly ( 19 ). Still, these synthetic approaches primarily rely on the same building blocks that biology already uses for assembling functional hierarchical structures (i.e., DNA and peptides) or rely on the elaborate design and fabrication of orthogonal units within synthetic organic ligands.…”