Self-Assembly of Temperature-Responsive Protein-Polymer Bioconjugates

Overview

Journal Bioconjug Chem

Publisher American Chemical Society

Specialty Biochemistry

Date 2015 Jun 18

PMID 26083370

Citations 15

Authors

Dafni Moatsou

Jian Li

Arnaz Ranji

Anais Pitto-Barry

Ioanna Ntai

Michael C Jewett

Rachel K OReilly

Affiliations

Soon will be listed here.

Abstract

We report a simple temperature-responsive bioconjugate system comprising superfolder green fluorescent protein (sfGFP) decorated with poly[(oligo ethylene glycol) methyl ether methacrylate] (PEGMA) polymers. We used amber suppression to site-specifically incorporate the non-canonical azide-functional amino acid p-azidophenylalanine (pAzF) into sfGFP at different positions. The azide moiety on modified sfGFP was then coupled using copper-catalyzed "click" chemistry with the alkyne terminus of a PEGMA synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization. The protein in the resulting bioconjugate was found to remain functionally active (i.e., fluorescent) after conjugation. Turbidity measurements revealed that the point of attachment of the polymer onto the protein scaffold has an impact on the thermoresponsive behavior of the resultant bioconjugate. Furthermore, small-angle X-ray scattering analysis showed the wrapping of the polymer around the protein in a temperature-dependent fashion. Our work demonstrates that standard genetic manipulation combined with an expanded genetic code provides an easy way to construct functional hybrid biomaterials where the location of the conjugation site on the protein plays an important role in determining material properties. We anticipate that our approach could be generalized for the synthesis of complex functional materials with precisely defined domain orientation, connectivity, and composition.

Citing Articles

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Supramolecular Assembly and Thermogelation Strategies Using Peptide-Polymer Conjugates.

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Peptide-Protein Coassemblies into Hierarchical and Bioactive Tubular Membranes.

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References

Velonia K, Rowan A, Nolte R . Lipase polystyrene giant amphiphiles. J Am Chem Soc. 2002; 124(16):4224-5. DOI: 10.1021/ja017809b. View

Roberts M, Bentley M, Harris J . Chemistry for peptide and protein PEGylation. Adv Drug Deliv Rev. 2002; 54(4):459-76. DOI: 10.1016/s0169-409x(02)00022-4. View

Rostovtsev V, Green L, Fokin V, Sharpless K . A stepwise huisgen cycloaddition process: copper(I)-catalyzed regioselective "ligation" of azides and terminal alkynes. Angew Chem Int Ed Engl. 2002; 41(14):2596-9. DOI: 10.1002/1521-3773(20020715)41:14<2596::AID-ANIE2596>3.0.CO;2-4. View

Krishna O, Kiick K . Protein- and peptide-modified synthetic polymeric biomaterials. Biopolymers. 2010; 94(1):32-48. PMC: 4437713. DOI: 10.1002/bip.21333. View

Stayton P, Shimoboji T, Long C, Chilkoti A, Chen G, Harris J . Control of protein-ligand recognition using a stimuli-responsive polymer. Nature. 1995; 378(6556):472-4. DOI: 10.1038/378472a0. View

Chin J . Expanding and reprogramming the genetic code of cells and animals. Annu Rev Biochem. 2014; 83:379-408. DOI: 10.1146/annurev-biochem-060713-035737. View

Boyer C, Bulmus V, Liu J, Davis T, Stenzel M, Barner-Kowollik C . Well-defined protein-polymer conjugates via in situ RAFT polymerization. J Am Chem Soc. 2007; 129(22):7145-54. DOI: 10.1021/ja070956a. View

Park T, Hoffman A . Synthesis and characterization of a soluble, temperature-sensitive polymer-conjugated enzyme. J Biomater Sci Polym Ed. 1993; 4(5):493-504. DOI: 10.1163/156856293x00159. View

Abuchowski A, van Es T, Palczuk N, DAVIS F . Alteration of immunological properties of bovine serum albumin by covalent attachment of polyethylene glycol. J Biol Chem. 1977; 252(11):3578-81. View

10.

Oshiba Y, Tamaki T, Ohashi H, Hirakawa H, Yamaguchi S, Nagamune T . Effect of length of molecular recognition moiety on enzymatic activity switching. J Biosci Bioeng. 2013; 116(4):433-7. DOI: 10.1016/j.jbiosc.2013.04.003. View

11.

Kolb H, Finn M, Sharpless K . Click Chemistry: Diverse Chemical Function from a Few Good Reactions. Angew Chem Int Ed Engl. 2001; 40(11):2004-2021. DOI: 10.1002/1521-3773(20010601)40:11<2004::AID-ANIE2004>3.0.CO;2-5. View

12.

Ding Z, Chen G, Hoffman A . Synthesis and purification of thermally sensitive oligomer-enzyme conjugates of poly(N-isopropylacrylamide)-trypsin. Bioconjug Chem. 1996; 7(1):121-6. DOI: 10.1021/bc950087r. View

13.

Canalle L, Lowik D, van Hest J . Polypeptide-polymer bioconjugates. Chem Soc Rev. 2009; 39(1):329-53. DOI: 10.1039/b807871h. View

14.

Averick S, Karacsony O, Mohin J, Yong X, Moellers N, Woodman B . Cooperative, reversible self-assembly of covalently pre-linked proteins into giant fibrous structures. Angew Chem Int Ed Engl. 2014; 53(31):8050-5. DOI: 10.1002/anie.201402827. View

15.

Hong S, Ntai I, Haimovich A, Kelleher N, Isaacs F, Jewett M . Cell-free protein synthesis from a release factor 1 deficient Escherichia coli activates efficient and multiple site-specific nonstandard amino acid incorporation. ACS Synth Biol. 2013; 3(6):398-409. PMC: 4065633. DOI: 10.1021/sb400140t. View

16.

Bulmus V, Ding Z, Long C, Stayton P, Hoffman A . Site-specific polymer-streptavidin bioconjugate for pH-controlled binding and triggered release of biotin. Bioconjug Chem. 2000; 11(1):78-83. DOI: 10.1021/bc9901043. View

17.

Young T, Ahmad I, Yin J, Schultz P . An enhanced system for unnatural amino acid mutagenesis in E. coli. J Mol Biol. 2009; 395(2):361-74. DOI: 10.1016/j.jmb.2009.10.030. View

18.

Hannink J, Cornelissen J, Farrera J, Foubert P, De Schryver F, Sommerdijk N . Protein-Polymer Hybrid Amphiphiles This research was supported by the Netherlands Foundation for Chemical Research (CW-NWO), the EC TMR Sisitomas and ESF Smarton programs, and the Ministerio de Educación y Cultura (Spain). The authors thank H. P. M..... Angew Chem Int Ed Engl. 2002; 40(24):4732-4734. View

19.

Ding Z, Chen G, Hoffman A . Unusual properties of thermally sensitive oligomer-enzyme conjugates of poly(N-isopropylacrylamide)-trypsin. J Biomed Mater Res. 1998; 39(3):498-505. DOI: 10.1002/(sici)1097-4636(19980305)39:3<498::aid-jbm22>3.0.co;2-5. View

20.

Abuchowski A, McCoy J, Palczuk N, van Es T, DAVIS F . Effect of covalent attachment of polyethylene glycol on immunogenicity and circulating life of bovine liver catalase. J Biol Chem. 1977; 252(11):3582-6. View