Effect of small molecule osmolytes on the self-assembly and functionality of globular protein-polymer diblock copolymers.
Blending the small molecule osmolytes glycerol and trehalose with the model globular protein-polymer block copolymer mCherry-b-poly(N-isopropyl acrylamide) (mCherry-b-PNIPAM) is demonstrated to improve protein functionality in self-assembled nanostructures. The incorporation of either additive into block copolymers results in functionality retention in the solid state of 80 and 100% for PNIPAM volume fractions of 40 and 55%, respectively. This represents a large improvement over the 50-60% functionality observed in the absence of any additive. Furthermore, glycerol decreases the thermal stability of block copolymer films by 15-20 °C, while trehalose results in an improvement in the thermal stability by 15-20 °C. These results suggest that hydrogen bond replacement is responsible for the retention of protein function but suppression or enhancement of thermal motion based on the glass transition of the osmolyte primarily determines thermal stability. While both osmolytes are observed to have a disordering effect on the nanostructure morphology with increasing concentration, this effect is less pronounced in materials with a larger polymer volume fraction. Glycerol preferentially localizes in the protein domains and swells the nanostructures, inducing disordering or a change in morphology depending on the PNIPAM coil fraction. In contrast, trehalose is observed to macrophase separate from the block copolymer, which results in nanodomains becoming more disordered without changing significantly in size.
Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.
Carla S
CS
Thomas
Liza
L
Xu
Bradley D
BD
Olsen
D000180:Acrylic Resins / Q000737:chemistry*; D005990:Glycerol / Q000737:chemistry*; D006860:Hydrogen Bonding; D008164:Luminescent Proteins / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry*; D009994:Osmolar Concentration; D010316:Particle Size; D055503:Protein Multimerization; D055550:Protein Stability; D053838:Scattering, Small Angle; D014199:Trehalose / Q000737:chemistry*; D014961:X-Ray Diffraction; D000097573:Red Fluorescent Protein
D000180:Acrylic Resins; D005990:Glycerol; D008164:Luminescent Proteins; D014199:Trehalose; D000097573:Red Fluorescent Protein