A Rationally Designed Supercharged Protein-Enzyme Chimera Self-Assembles In Situ to Yield Bifunctional Composite Textiles

Abstract

Catalytically active materials for the enhancement of personalized protective equipment (PPE) could be advantageous to help alleviate threats posed by neurotoxic organophosphorus compounds (OPs). Accordingly, a chimeric protein comprised of a supercharged green fluorescent protein (scGFP) and phosphotriesterase from Agrobacterium radiobacter (arPTE) was designed to drive the polymer surfactant (S − )-mediated selfassembly of microclusters to produce robust, enzymatically active materials. The chimera scGFP-arPTE was structurally characterized via circular dichroism spectroscopy and synchrotron radiation small-angle X-ray scattering, and its biophysical properties were determined. Significantly, the chimera exhibited greater thermal stability than the native constituent proteins, as well as a higher catalytic turnover number (k cat ). Furthermore, scGFP-arPTE was electrostatically complexed with monomeric S − , driving selfassembly into [scGFP-arPTE][S − ] nanoclusters, which could be dehydrated and cross-linked to yield enzymatically active [scGFP-arPTE][S − ] porous films with a high-order structure. Moreover, these clusters could self-assemble within cotton fibers to generate active composite textiles without the need for the pretreatment of the fabrics. Significantly, the resulting materials maintained the biophysical activities of both constituent proteins and displayed recyclable and persistent activity against the nerve agent simulant paraoxon.

Modular Bioorthogonal Lipid Nanoparticle Modification Platforms for Cardiac Homing

“…The membrane anchor domain (protein fusion 1 ) showed a 10% increase in the radius of gyration ( R g ) upon surfactant conjugation (from 27.4 ± 0.6 to 30.1 ± 0.6 Å), which was accompanied by a 5.5 Å increase (133.5 to 139 Å) in the D max (maximum distance between two atoms), which was calculated from the pair-distance distribution P (r) (Figures b,c and S10). These data indicated that the surfactant electrostatically bound to the anchor domain to form a compact polymer surfactant corona . The Spy-tagged coupling partners CshA-mCh-ST3 (protein fusion 2 ) and mCh-ST3 (protein fusion 3 ) had R g ’s of 58 ± 3 nm and 21.48 ± 0.09 Å and D max values of 239.5 and 95.5 Å, respectively, which demonstrated that the large intrinsically disordered domain of CshA maintained an extended configuration …”

Carrier Variety Used in Immobilization of His6-OPH Extends Its Application Areas

“…is based that fast and highly effective immobilization, and then isolation and purification of the enzyme itself occurs. The emerging coordination bonds between metal atoms in the carrier and nitrogen atoms in the imidazole rings of the polyhistidine sequence in the protein molecule make it possible to anchor the enzyme on the carrier quite firmly [ 2 , 22 , 23 ] and use it for a long time, in such an immobilized form, for the destruction of OPCs ( Table 1 , [ 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 ]). Of particular interest are such immobilization variants for the stabilization and repeated use of the enzyme in the degradation of OPC-polluting water systems [ 22 , 23 ].…”

“…A fundamentally different approach, based on the use of polyelectrolyte interactions of the surface of the His-tagged enzyme with variously charged surfactants, was used by researchers for the creation of various bioconjugates to improve the stability and catalytic characteristics of the enzyme [ 33 , 34 , 35 ]. At the same time, the expediency of using such coatings for the enzyme, including forming a capsule in the form of a so-called corona, was noted.…”

“…This creates scientific and practical foundations for the development of personal protective equipment against the toxic effects of OPCs. Additional genetic modifications of the enzyme itself—particularly due to the connection with the fusion protein (GFP) [ 33 ] before its encapsulation in the layers of surfactants—create the possibility of additional control of the effect of immobilization conditions on the properties of the main biocatalyst through the properties of the selected fusion partner (fluorescence intensity of GFP).…”

“…It should be emphasized that the catalytic activity was studied against different substrates (methyl parathion [ 23 , 25 , 26 , 27 , 28 , 29 , 30 , 37 , 46 , 47 ], paraoxon [ 31 , 32 , 33 , 34 , 41 , 42 , 43 , 44 , 46 , 47 ], malathion [ 36 , 44 ], methyl–paraoxon [ 38 ], coumaphos [ 39 , 40 ], chlorphyryphos [ 45 ], VX [ 8 , 29 ], etc.) under different conditions (mixing and its absence [ 25 , 32 , 44 , 45 ], flow reactor [ 23 , 82 ]) at different temperatures and pH values [ 44 ].…”