Recombinant production of self-assembling β-structured peptides using SUMO as a fusion partner

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• 作者：Abhinav Prakash (1)
  Stephen J Parsons (1)
  Stuart Kyle (1) (2)
  Michael J McPherson (1) (2)
  
• 关键词：Self ; assembly ; Peptide ; Hydrogel ; Recombinant expression ; Scaffold
• 刊名：Microbial Cell Factories
• 出版年：2012
• 出版时间：December 2012
• 年：2012
• 卷：11
• 期：1
• 全文大小：552KB
• 参考文献：1. Webber MJ, Tongers J, Newcomb CJ, Marquardt KT, Bauersachs J, Losordo DW, Stupp SI: Supramolecular nanostructures that mimic VEGF as a strategy for ischemic tissue repair. / Proc Natl Acad Sci USA 2011, 108:13438-3443. CrossRef
  2. Cho H, Balaji S, Sheikh AQ, Hurley JR, Tian YF, Collier JH, Crombleholme TM, Narmoneva DA: Regulation of endothelial cell activation and angiogenesis by injectable peptide nanofibers. / Acta Biomater 2012, 8:154-64. CrossRef
  3. Gelain F, Unsworth LD, Zhang S: Slow and sustained release of active cytokines from self-assembling peptide scaffolds. / J Control Release 2010, 145:231-39. CrossRef
  4. Bell CJ, Carrick LM, Katta J, Jin Z, Ingham E, Aggeli A, Boden N, Waigh TA, Fisher J: Self-assembling peptides as injectable lubricants for osteoarthritis. / J Biomed Mater Res A 2006, 78:236-46.
  5. Firth A, Aggeli A, Burke JL, Yang XB, Kirkham J: Biomimetic self-assembling peptides as injectable scaffolds for hard tissue engineering. / Nanomedicine 2006, 1:189-99. CrossRef
  6. Kirkham J, Firth A, Vernals D, Boden N, Robinson C, Shore RC, Brookes SJ, Aggeli A: Self-assembling peptide scaffolds promote enamel remineralization. / J Dent Res 2007, 86:426-30. CrossRef
  7. Aggeli A, Bell M, Boden N, Keen JN, Knowles PF, McLeish TCB, Pitkeathly M, Radford SE: Responsive gels formed by the spontaneous self-assembly of peptides into polymeric beta-sheet tapes. / Nature 1997, 386:259-62. CrossRef
  8. Aggeli A, Bell M, Boden N, Carrick LM, Strong AE: Self-assembling peptide polyelectrolyte beta-sheet complexes form nematic hydrogels. / Angew Chemie Int Ed Engl 2003, 42:5603-606. CrossRef
  9. Aggeli A, Nyrkova IA, Bell M, Harding R, Carrick L, McLeish TCB, Semenov AN, Boden N: Hierarchical self-assembly of chiral rod-like molecules as a model for peptide beta-sheet tapes, ribbons, fibrils, and fibers. / Proc Natl Acad Sci USA 2001, 98:11857-1862. CrossRef
  10. Kyle S, Aggeli A, Ingham E, McPherson MJ: Recombinant self-assembling peptides as biomaterials for tissue engineering. / Biomaterials 2010, 31:9395-405. CrossRef
  11. Kyle S, McPherson MJ, Aggeli A, Ingham E: Rational molecular design of complementary self-assembling peptide hydrogels. / Adv Healthcare Mater in press
  12. Riley JM, Aggeli A, Koopmans RJ, McPherson MJ: Bioproduction and characterization of a pH responsive self-assembling peptide. / Biotechnol Bioeng 2009, 103:241-51. CrossRef
  13. Kyle S, Aggeli A, Ingham E, McPherson MJ: Production of self-assembling biomaterials for tissue engineering. / Trends Biotechnol 2009, 27:423-33. CrossRef
  14. McPherson MJ, James K, Kyle S, Parsons S, Riley J: Recombinant production of self-assembling peptides. In / Engineering aspects of self-organising materials. Volume 35. Koopmans RJ: Elsevier; 2009:80-17.
  15. Hartmann BM, Kaar W, Falconer RJ, Zeng B, Middelberg APJ: Expression and purification of a nanostructure-forming peptide. / J Biotechnol 2008, 135:85-1. CrossRef
  16. Studier FW: Protein production by auto-induction in high-density shaking cultures. / Protein Expres Purif 2005, 41:207-34. CrossRef
  17. Hartmann BM, Kaar W, Yoo IK, Lua LH, Falconer RJ, Middelberg AJP: The chromatography-free release, isolation and purification of recombinant peptide for fibril self-assembly. / Biotechnol Bioeng 2009, 104:973-85. CrossRef
  18. Satakarni M, Curtis R: Production of recombinant peptides as fusions with SUMO. / Protein Expres Purif 2011, 78:113-19. CrossRef
  19. Malakhov M, Mattern M, Malakhova O, Drinker M, Weeks S, Butt T: SUMO fusions and SUMO-specific protease for efficient expression and purification of proteins. / J Struct Funct Genomics 2004, 5:75-6. CrossRef
  20. Lee CD, Sun HC, Hu SM, Chiu CF, Homhuan A, Liang SM, Leng CH, Wang TF: An improved SUMO fusion protein system for effective production of native proteins. / Protein Sci 2008, 17:1241-248. CrossRef
  21. van Hell AJ, Costa C, Flesch FM, Sutter M, Jiskoot W, Crommelin DJA, Hennink WE, Mastrobattista E: Self-assembly of recombinant amphiphilic oligopeptides into vesicles. / Biomacromolecules 2007, 8:2753-761. CrossRef
  22. Satakarni M, Koutinas AA, Webb C, Curtis R: Enrichment of fermentation media and optimization of expression conditions for the production of EAK16 peptide as fusions with SUMO. / Biotechnol Bioeng 2009, 102:725-35. CrossRef
  23. Bosse-Doenecke E, Weininger U, Gopalswamy M, Balbach J, Knudsen SM, Rudolph R: High yield production of recombinant native and modified peptides exemplified by ligands for G-protein coupled receptors. / Protein Expres Purif 2008, 58:114-21. CrossRef
  24. Butt TR, Edavettal SC, Hall JP, Mattern MR: SUMO fusion technology for difficult-to-express proteins. / Protein Expres Purif 2005, 43:1-. CrossRef
  25. Marblestone JG, Edavettal SC, Lim Y, Lim P, Zuo X, Butt TR: Comparison of SUMO fusion technology with traditional gene fusion systems: Enhanced expression and solubility with SUMO. / Protein Sci 2006, 15:182-89. CrossRef
  26. Zuo X, Li S, Hall J, Mattern M, Tran H, Shoo J, Tan R, Weiss SR, Butt TR: Enhanced expression and purification of membrane proteins by SUMO fusion in Escherichia coli. / J Struct Funct Genomics 2005, 6:103-11. CrossRef
  27. Guzzo CM, Yang DCH: Systematic analysis of fusion and affinity tags using human aspartyl-tRNA synthetase expressed in E. coli. / Protein Expres Purif 2007, 54:166-75. CrossRef
  28. Li J, Zhang J, Zhang Z, Ma H, Zhang J, Zhang S: Production of bioactive human beta-defensin-4 in Escherichia coli using SUMO fusion partner. / Protein J 2010, 29:314-19. CrossRef
  29. Sambrook J: / Fritsch EF, Maniatis T: Molecular cloning: A laboratory manual. New York: Cold Spring Harbor Laboratory Press, Cold Spring Harbor; 1989.
  30. Assenberg R, Delmas O, Graham SC, Verma A, Berrow N, Stuart DI, Owens RJ, Bourhy H, Grimes JM: Expression, purification and crystallization of a lyssavirus matrix (M) protein. / Acta Cryst 2008, F64:258-62.
  
• 作者单位：Abhinav Prakash (1)
  Stephen J Parsons (1)
  Stuart Kyle (1) (2)
  Michael J McPherson (1) (2)
  
  1. Institute of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, LS2 9JT, UK
  2. Astbury Centre for Structural Molecular Biology Faculty of Biological Sciences, University of Leeds, Leeds, LS2 9JT, UK
  
• ISSN：1475-2859

文摘

Background Self-assembling peptides that form nanostructured hydrogels are important biomaterials for tissue engineering scaffolds. The P11-family of peptides includes, P11-4 (QQRFEWEFEQQ) and the complementary peptides P11-13 (EQEFEWEFEQE) and P11-14 (QQOrnFOrnWOrnFOrnQQ). These form self-supporting hydrogels under physiological conditions (pH 7.4, 140 mM NaCl) either alone (P11-4) or when mixed (P11-13 and P11-14). We report a SUMO-peptide expression strategy suitable for allowing release of native sequence peptide by SUMO protease cleavage. Results We have expressed SUMO-peptide fusion proteins from pET vectors by using autoinduction methods. Immobilised metal affinity chromatography was used to purify the fusion protein, followed by SUMO protease cleavage in water to release the peptides, which were recovered by reverse phase HPLC. The peptide samples were analysed by electrospray mass spectrometry and self-assembly was followed by circular dichroism and transmission electron microscopy. Conclusions The fusion proteins were produced in high yields and the β-structured peptides were efficiently released by SUMO protease resulting in peptides with no additional amino acid residues and with recoveries of 46% to 99%. The peptides behaved essentially the same as chemically synthesised and previously characterised recombinant peptides in self-assembly and biophysical assays.