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      Polyproline as a Minimal Antifreeze Protein Mimic That Enhances the Cryopreservation of Cell Monolayers

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          Abstract

          Tissue engineering, gene therapy, drug screening, and emerging regenerative medicine therapies are fundamentally reliant on high‐quality adherent cell culture, but current methods to cryopreserve cells in this format can give low cell yields and require large volumes of solvent “antifreezes”. Herein, we report polyproline as a minimum (bio)synthetic mimic of antifreeze proteins that is accessible by solution, solid‐phase, and recombinant methods. We demonstrate that polyproline has ice recrystallisation inhibition activity linked to its amphipathic helix and that it enhances the DMSO cryopreservation of adherent cell lines. Polyproline may be a versatile additive in the emerging field of macromolecular cryoprotectants.

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          Most cited references53

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          Solid Phase Peptide Synthesis. I. The Synthesis of a Tetrapeptide

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            Biopharmaceutical benchmarks 2014.

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              eIF5A promotes translation of polyproline motifs.

              Translation factor eIF5A, containing the unique amino acid hypusine, was originally shown to stimulate Met-puromycin synthesis, a model assay for peptide bond formation. More recently, eIF5A was shown to promote translation elongation; however, its precise requirement in protein synthesis remains elusive. We use in vivo assays in yeast and in vitro reconstituted translation assays to reveal a specific requirement for eIF5A to promote peptide bond formation between consecutive Pro residues. Addition of eIF5A relieves ribosomal stalling during translation of three consecutive Pro residues in vitro, and loss of eIF5A function impairs translation of polyproline-containing proteins in vivo. Hydroxyl radical probing experiments localized eIF5A near the E site of the ribosome with its hypusine residue adjacent to the acceptor stem of the P site tRNA. Thus, eIF5A, like its bacterial ortholog EFP, is proposed to stimulate the peptidyl transferase activity of the ribosome and facilitate the reactivity of poor substrates like Pro. Copyright © 2013 Elsevier Inc. All rights reserved.
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                Author and article information

                Contributors
                m.i.gibson@warwick.ac.uk
                Journal
                Angew Chem Int Ed Engl
                Angew. Chem. Int. Ed. Engl
                10.1002/(ISSN)1521-3773
                ANIE
                Angewandte Chemie (International Ed. in English)
                John Wiley and Sons Inc. (Hoboken )
                1433-7851
                1521-3773
                22 November 2017
                11 December 2017
                : 56
                : 50 ( doiID: 10.1002/anie.v56.50 )
                : 15941-15944
                Affiliations
                [ 1 ] Department of Chemistry University of Warwick Gibbet Hill Road Coventry CV47 AL UK
                [ 2 ] Warwick Medical School University of Warwick Coventry CV4 7AL UK
                [ 3 ] Ceramics Synthesis and Functionalization Lab UMR3080 CNRS/Saint-Gobain 550 Avenue Alphonse Jauffret 84306 Cavaillon France
                Author information
                http://orcid.org/0000-0003-1313-6874
                http://orcid.org/0000-0002-5448-1722
                http://orcid.org/0000-0002-9569-6738
                http://orcid.org/0000-0002-3363-3184
                http://orcid.org/0000-0002-8297-1278
                Article
                ANIE201706703
                10.1002/anie.201706703
                5722203
                29044869
                1a23ec7c-d8e7-4663-805f-fc8b5f4dd1fd
                © 2017 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

                This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 04 July 2017
                : 27 September 2017
                Page count
                Figures: 6, Tables: 1, References: 41, Pages: 4, Words: 0
                Funding
                Funded by: H2020 European Research Council
                Award ID: 638661
                Award ID: 278004
                Funded by: Engineering and Physical Sciences Research Council
                Award ID: EP/F500378/1
                Categories
                Communication
                Communications
                Biomaterials
                Custom metadata
                2.0
                anie201706703
                December 11, 2017
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.2.8 mode:remove_FC converted:12.01.2018

                Chemistry
                biomaterials,cryopreservation,ice recrystallization inhibitors,monolayers,polymers
                Chemistry
                biomaterials, cryopreservation, ice recrystallization inhibitors, monolayers, polymers

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