Random heteropolymers preserve protein function in foreign environments
View Patent ↗Compositions comprise statistically random heteropolymers complexed with active proteins, and are formulated and used in stimuli-responsive materials and nanoreactors composed of proteins and synthetic materials.
1. A method of preserving protein function in foreign environments, the method comprising forming a composition comprising a complex of an active protein and statistically random heteropolymers (SRHPs) in an organic solvent, and detecting activity of the protein in the composition, wherein the SRHPs comprising monomers: methyl methacrylate (MMA), oligo(ethylene glycol) methacrylate (OEGMA), 3-sulfopropyl methacrylate potassium salt (3-SPMA) and 2-ethylhexyl methacrylate (2-EHMA).
2. The method of claim 1 , wherein the heteropolymers disperse in both aqueous and organic media.
3. The method of claim 1 , wherein a distribution histogram of monomer blocks of the heteropolymers decrease in normalized frequency from block size 1, wherein block size 10 has a normalized frequency of less than 1%, and block size 1 has a normalized frequency of 5-20%.
4. The method of claim 1 , wherein the protein is an enzyme or fluorescent protein.
5. The method of claim 1 , wherein the solvent is selected from 2-propanol, acetone, acetonitrile, chloroform, dichloromethane, dimethyl sulfoxide, ethyl acetate, hexane, methanol, tetrahydrofuran, and toluene.
6. The method of claim 1 , wherein the SRHPs comprising monomers: methyl methacrylate (MMA), oligo(ethylene glycol) methacrylate (OEGMA), 3-sulfopropyl methacrylate potassium salt (3-SPMA) and 2-ethylhexyl methacrylate (2-EHMA), in ratio: 5(MMA):2.5(OEGMA):2(2-EHMA):0.5(3-SPMA).
7. The method of claim 1 , wherein:
the protein is an enzyme or fluorescent protein;
the solvent is selected from 2-propanol, acetone, acetonitrile, chloroform, dichloromethane, dimethyl sulfoxide, ethyl acetate, hexane, methanol, tetrahydrofuran, and toluene; and
the SRHPs comprising varying ratios a plurality of monomers selected from methyl methacrylate (MMA), oligo(ethylene glycol) methacrylate (OEGMA), 3-sulfopropyl methacrylate potassium salt (3-SPMA) and 2-ethylhexyl methacrylate (2-EHMA).
8. The method of claim 1 , wherein:
the heteropolymers disperse in both aqueous and organic media;
a distribution histogram of monomer blocks of the heteropolymers decrease in normalized frequency from block size 1, wherein block size 10 has a normalized frequency of less than 1%, and block size 1 has a normalized frequency of 5-20%;
the protein is an enzyme or fluorescent protein;
the solvent is selected from 2-propanol, acetone, acetonitrile, chloroform, dichloromethane, dimethyl sulfoxide, ethyl acetate, hexane, methanol, tetrahydrofuran, and toluene; and
the SRHPs comprising varying ratios a plurality of monomers selected from methyl methacrylate (MMA), oligo(ethylene glycol) methacrylate (OEGMA), 3-sulfopropyl methacrylate potassium salt (3-SPMA) and 2-ethylhexyl methacrylate (2-EHMA).
9. The method of claim 1 , wherein the protein is organophosphorus hydrolase (OPH).
10. The method of claim 1 , wherein the protein and SRHPs are contained in a polymer matrix.
11. The method of claim 1 , wherein the protein is organophosphorus hydrolase (OPH), and the protein and SRHPs are contained in a polymer matrix, and the matrix comprises polymerized polyethylene oxide (PEO) or PMMA in the form of a fiber mat.
12. The method of claim 6 , wherein the protein is organophosphorus hydrolase (OPH), and the protein and SRHPs are contained in a polymer matrix, and the matrix comprises polymerized polyethylene oxide (PEO) or PMMA in the form of a fiber mat.
13. The method of claim 1 , wherein the forming step comprises mixing the protein and the SRHPs in an aqueous solution; drying the mixture; resuspending the dried mixture in the organic solvent, forming the composition.
14. The method of claim 1 , wherein the forming step is in a protein expression system, wherein the protein is being expressed and incorporated in the complex, wherein the complex enhances activity of the protein, compared an expression system without the SRHPs.
15. The method of claim 1 , wherein the forming step is in a protein expression system, wherein the protein is being expressed and incorporated in the complex, wherein the complex enhances activity of the protein, compared an expression system without the SRHPs and the expression system is a cell-free system.
16. The method of claim 1 , further comprising contacting the complex with a liposome under conditions wherein the protein relocates from the complex to the liposome.
17. The method of claim 1 , further comprising contacting the complex with a liposome under conditions wherein the protein relocates from the complex to the liposome, in form of an active, transmembrane protein.