Tuning the Properties of Protein-Based Polymers Using High-Performance Orthogonal Translation Systems for the Incorporation of Aromatic Non-Canonical Amino Acids.
elastin-like polypeptides (ELP)
genetic code expansion
non-canonical amino acids (ncAAs)
resilin-like polypeptides (RLP)
smart biomaterials
Journal
Frontiers in bioengineering and biotechnology
ISSN: 2296-4185
Titre abrégé: Front Bioeng Biotechnol
Pays: Switzerland
ID NLM: 101632513
Informations de publication
Date de publication:
2022
2022
Historique:
received:
05
04
2022
accepted:
27
04
2022
entrez:
17
6
2022
pubmed:
18
6
2022
medline:
18
6
2022
Statut:
epublish
Résumé
The incorporation of non-canonical amino acids (ncAAs) using engineered aminoacyl-tRNA synthetases (aaRSs) has emerged as a powerful methodology to expand the chemical repertoire of proteins. However, the low efficiencies of typical aaRS variants limit the incorporation of ncAAs to only one or a few sites within a protein chain, hindering the design of protein-based polymers (PBPs) in which multi-site ncAA incorporation can be used to impart new properties and functions. Here, we determined the substrate specificities of 11 recently developed high-performance aaRS variants and identified those that enable an efficient multi-site incorporation of 15 different aromatic ncAAs. We used these aaRS variants to produce libraries of two temperature-responsive PBPs-elastin- and resilin-like polypeptides (ELPs and RLPs, respectively)-that bear multiple instances of each ncAA. We show that incorporating such aromatic ncAAs into the protein structure of ELPs and RLPs can affect their temperature responsiveness, secondary structure, and self-assembly propensity, yielding new and diverse families of ELPs and RLPs, each from a single DNA template. Finally, using a molecular model, we demonstrate that the temperature-responsive behavior of RLPs is strongly affected by both the hydrophobicity and the size of the unnatural aromatic side-chain. The ability to efficiently incorporate multiple instances of diverse ncAAs alongside the 20 natural amino acids can help to elucidate the effect of ncAA incorporation on these and many other PBPs, with the aim of designing additional precise and chemically diverse polymers with new or improved properties.
Identifiants
pubmed: 35711629
doi: 10.3389/fbioe.2022.913057
pii: 913057
pmc: PMC9195583
doi:
Types de publication
Journal Article
Langues
eng
Pagination
913057Informations de copyright
Copyright © 2022 Gueta, Sheinenzon, Azulay, Shalit, Strugach, Hadar, Gelkop, Milo and Amiram.
Déclaration de conflit d'intérêts
MA, DH, DSS, and SG filed a patent related to this technology. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Références
Proc Natl Acad Sci U S A. 2022 Jan 25;119(4):
pubmed: 35046019
Biopolymers. 2010;94(1):60-77
pubmed: 20091871
Biochim Biophys Acta. 2014 Jun;1844(6):1059-70
pubmed: 24631543
Polym Chem. 2014 Jan;5(5):1614-1625
pubmed: 24511327
J Am Chem Soc. 2008 Mar 26;130(12):4028-33
pubmed: 18321101
Bioconjug Chem. 2018 Jun 20;29(6):1876-1884
pubmed: 29786419
Curr Opin Biotechnol. 2019 Dec;60:168-178
pubmed: 30974337
Nat Struct Biol. 2003 Jun;10(6):425-32
pubmed: 12754495
ACS Macro Lett. 2019 Dec 17;8(12):1648-1653
pubmed: 35619386
Proc Natl Acad Sci U S A. 2020 May 26;117(21):11421-11431
pubmed: 32393642
Mol Cells. 2019 May 31;42(5):386-396
pubmed: 31122001
Nat Commun. 2021 Jan 8;12(1):149
pubmed: 33420053
Biomacromolecules. 2013 Apr 8;14(4):976-85
pubmed: 23406497
Nature. 2014 Mar 13;507(7491):210-4
pubmed: 24622199
Angew Chem Int Ed Engl. 2006 Dec 11;45(48):8141-5
pubmed: 17109457
J Am Chem Soc. 2005 Dec 28;127(51):18121-32
pubmed: 16366565
Mol Cell. 2015 Mar 5;57(5):936-947
pubmed: 25747659
Biochemistry. 2018 May 1;57(17):2478-2487
pubmed: 29517898
Adv Drug Deliv Rev. 2020;156:133-187
pubmed: 32871201
Structure. 2009 Mar 11;17(3):335-44
pubmed: 19278648
J Control Release. 2013 Nov 28;172(1):144-151
pubmed: 23928357
Org Biomol Chem. 2021 Aug 28;19(32):7031-7040
pubmed: 34333582
Q Rev Biophys. 2017 Jan;50:e7
pubmed: 29233219
Chembiochem. 2017 Sep 19;18(18):1819-1823
pubmed: 28650092
Nano Lett. 2019 Jan 9;19(1):247-254
pubmed: 30540482
Molecules. 2018 Jul 08;23(7):
pubmed: 29986538
Synth Syst Biotechnol. 2018 Oct 03;3(3):150-158
pubmed: 30345400
Angew Chem Int Ed Engl. 2018 Oct 26;57(44):14350-14361
pubmed: 30144241
Nature. 2017 Oct 4;550(7674):53-60
pubmed: 28980641
Curr Opin Chem Biol. 2021 Aug;63:123-131
pubmed: 33845403
Biomacromolecules. 2002 Mar-Apr;3(2):357-67
pubmed: 11888323
Nat Chem. 2020 Sep;12(9):814-825
pubmed: 32747754
Polymers (Basel). 2021 May 01;13(9):
pubmed: 34062852
J Control Release. 2014 Sep 28;190:314-30
pubmed: 24979207
Curr Opin Chem Biol. 2010 Dec;14(6):774-80
pubmed: 21071259
Biomacromolecules. 2004 May-Jun;5(3):846-51
pubmed: 15132671
Biochemistry. 2011 Mar 22;50(11):1894-900
pubmed: 21280675
Proc Natl Acad Sci U S A. 2017 Sep 26;114(39):E8194-E8203
pubmed: 28894006
Biomacromolecules. 2013 Aug 12;14(8):2866-72
pubmed: 23808597
Curr Opin Chem Biol. 2018 Oct;46:115-122
pubmed: 30059834
Mol Cell. 2016 Jul 7;63(1):72-85
pubmed: 27392146
Biomacromolecules. 2019 Mar 11;20(3):1178-1189
pubmed: 30715857
Int J Mol Sci. 2019 May 11;20(9):
pubmed: 31083552
Cell. 2018 Jul 26;174(3):688-699.e16
pubmed: 29961577
Chembiochem. 2021 Jan 5;22(1):120-123
pubmed: 32815262
Biopolymers. 1992 Sep;32(9):1243-50
pubmed: 1420991
Protein Sci. 2017 Sep;26(9):1785-1795
pubmed: 28639381
Sci Rep. 2015 May 18;5:9762
pubmed: 25985257
ACS Chem Biol. 2018 Apr 20;13(4):854-870
pubmed: 29345901
Science. 2013 Oct 18;342(6156):357-60
pubmed: 24136966
J Am Chem Soc. 2008 Jan 16;130(2):687-94
pubmed: 18085778
Nat Chem. 2018 Aug;10(8):831-837
pubmed: 29807989
Nat Biotechnol. 2015 Dec;33(12):1272-1279
pubmed: 26571098
J Mol Biol. 2018 Nov 2;430(23):4619-4635
pubmed: 29949750
Curr Protoc Protein Sci. 2010 Aug;Chapter 6:Unit 6.11
pubmed: 20814933
Macromol Biosci. 2021 Sep;21(9):e2100129
pubmed: 34145967
J Control Release. 2016 Oct 28;240:93-108
pubmed: 26578439
Curr Opin Biotechnol. 2017 Dec;48:1-7
pubmed: 28237511
Biomacromolecules. 2017 Feb 13;18(2):544-550
pubmed: 28075561
ACS Macro Lett. 2015 Nov 17;4(11):1283-1286
pubmed: 35614829
Chembiochem. 2021 Apr 16;22(8):1379-1384
pubmed: 33350556
J Biol Chem. 2017 Nov 17;292(46):19110-19120
pubmed: 28924037
Nat Chem Biol. 2013 Oct;9(10):594-8
pubmed: 24045798
Mol Syst Des Eng. 2020 Aug 1;5(7):1239-1254
pubmed: 33796336