Ukr.Biochem.J. 2018; Volume 90, Issue 1, Jan-Feb, pp. 68-76

doi: https://doi.org/10.15407/ubj90.01.068

Designing, docking and heterologous expression of an anti-HER2 affibody molecule

N. Salmanian Tabasi1, A. Gholizadeh1, B. Baghban Kohnehrouz2

1Research Institute for Fundamental Sciences, University of Tabriz, Iran;
2Department of Plant Breeding and Biotechnology, University of Tabriz, Iran;
e-mail: aghz_bioch@yahoo.co.in

Affibody molecules are small protein scaffolds mostly based on triple-helical bundle protein domains. Many triple helix-based affibody proteins have shown prominent properties for tumor imaging and therapy. In our opinion, the structural organizations and the sizes of affibody molecules could be modified to increase their recognition abilities and binding affinities to human epidermal growth factor receptor type 2 (HER2). Thereby, the purpose of this study was to design and characterize a novel platform of affibody molecule consisting of five separate helixes (encoding 99 amino acids with a duplicate of helixes 1 and 2 at N-terminus plus GGGC chelator peptide sequence at C-terminus) enable of binding to HER2 with higher avidity. Using in silico screening methods, the structure and the interactive potential of designed affibody was comparatively investigated. The molecular expression and production of the designed affibody in Escherichia coli cells was successfully examined and reported.

 

Keywords: , , ,


References:

  1. Feldwisch J, Tolmachev V, Lendel C, Herne N, Sjöberg A, Larsson B, Rosik D, Lindqvist E, Fant G, Höidén-Guthenberg I, Galli J, Jonasson P, Abrahmsén L. Design of an optimized scaffold for affibody molecules. J Mol Biol. 2010 Apr 30;398(2):232-47. PubMed, CrossRef
  2. Löfblom J, Feldwisch J, Tolmachev V, Carlsson J, Ståhl S, Frejd FY. Affibody molecules: engineered proteins for therapeutic, diagnostic and biotechnological applications. FEBS Lett. 2010 Jun 18;584(12):2670-80. PubMed, CrossRef
  3. Nord K, Nilsson J, Nilsson B, Uhlén M, Nygren PA. A combinatorial library of an alpha-helical bacterial receptor domain. Protein Eng. 1995 Jun;8(6):601-8. PubMed, CrossRef
  4. Honarvar H, Jokilaakso N, Andersson K, Malmberg J, Rosik D, Orlova A, Karlström AE, Tolmachev V, Järver P. Evaluation of backbone-cyclized HER2-binding 2-helix affibody molecule for in vivo molecular imaging. Nucl Med Biol. 2013 Apr;40(3):378-86. PubMed, CrossRef
  5. Kronqvist N, Löfblom J, Jonsson A, Wernérus H, Ståhl S. A novel affinity protein selection system based on staphylococcal cell surface display and flow cytometry. Protein Eng Des Sel. 2008 Apr;21(4):247-55. PubMed, CrossRef
  6. Jonsson A, Wållberg H, Herne N, Ståhl S, Frejd FY. Generation of tumour-necrosis-factor-alpha-specific affibody molecules capable of blocking receptor binding in vitro. Biotechnol Appl Biochem. 2009 Aug 17;54(2):93-103. PubMed, CrossRef
  7. Wikman M, Steffen AC, Gunneriusson E, Tolmachev V, Adams GP, Carlsson J, Ståhl S. Selection and characterization of HER2/neu-binding affibody ligands. Protein Eng Des Sel. 2004 May;17(5):455-62. PubMed, CrossRef
  8. Orlova A, Magnusson M, Eriksson TL, Nilsson M, Larsson B, Höidén-Guthenberg I, Widström C, Carlsson J, Tolmachev V, Ståhl S, Nilsson FY. Tumor imaging using a picomolar affinity HER2 binding affibody molecule. Cancer Res. 2006 Apr 15;66(8):4339-48. PubMed, CrossRef
  9. Friedman M, Orlova A, Johansson E, Eriksson TL, Höidén-Guthenberg I, Tolmachev V, Nilsson FY, Ståhl S. Directed evolution to low nanomolar affinity of a tumor-targeting epidermal growth factor receptor-binding affibody molecule. J Mol Biol. 2008 Mar 7;376(5):1388-402.  PubMed, CrossRef
  10. Tolmachev V, Rosik D, Wållberg H, Sjöberg A, Sandström M, Hansson M, Wennborg A, Orlova A. Imaging of EGFR expression in murine xenografts using site-specifically labelled anti-EGFR 111In-DOTA-Z EGFR:2377 Affibody molecule: aspect of the injected tracer amount. Eur J Nucl Med Mol Imaging. 2010 Mar;37(3):613-22. PubMed, CrossRef
  11. Altai M, Wållberg H, Orlova A, Rosestedt M, Hosseinimehr SJ, Tolmachev V, Ståhl S. Order of amino acids in C-terminal cysteine-containing peptide-based chelators influences cellular processing and biodistribution of 99mTc-labeled recombinant Affibody molecules. Amino Acids. 2012 May;42(5):1975-85. PubMed, CrossRef
  12. Citri A, Yarden Y. EGF-ERBB signalling: towards the systems level. Nat Rev Mol Cell Biol. 2006 Jul;7(7):505-16. PubMed, CrossRef
  13. Hynes NE, Lane HA. ERBB receptors and cancer: the complexity of targeted inhibitors. Nat Rev Cancer. 2005 May;5(5):341-54. PubMed, CrossRef
  14. Tran TA, Rosik D, Abrahmsén L, Sandström M, Sjöberg A, Wållberg H, Ahlgren S, Orlova A, Tolmachev V. Design, synthesis and biological evaluation of a multifunctional HER2-specific Affibody molecule for molecular imaging. Eur J Nucl Med Mol Imaging. 2009 Nov;36(11):1864-73.  PubMed, CrossRef
  15. Xue X, Wang B, Du W, Zhang C, Song Y, Cai Y, Cen D, Wang L, Xiong Y, Jiang P, Zhu S, Zhao KN, Zhang L. Generation of affibody molecules specific for HPV16 E7 recognition. Oncotarget. 2016 Nov 8;7(45):73995-74005. PubMed, PubMedCentral, CrossRef
  16. Ahlgren S, Orlova A, Rosik D, Sandström M, Sjöberg A, Baastrup B, Widmark O, Fant G, Feldwisch J, Tolmachev V. Evaluation of maleimide derivative of DOTA for site-specific labeling of recombinant affibody molecules. Bioconjug Chem. 2008 Jan;19(1):235-43. PubMed, CrossRef
  17. Ren G, Zhang R, Liu Z, Webster JM, Miao Z, Gambhir SS, Syud FA, Cheng Z. A 2-helix small protein labeled with 68Ga for PET imaging of HER2 expression. J Nucl Med. 2009 Sep;50(9):1492-9. PubMed, PubMedCentral, CrossRef
  18. Lyakhov I, Zielinski R, Kuban M, Kramer-Marek G, Fisher R, Chertov O, Bindu L, Capala J. HER2- and EGFR-specific affiprobes: novel recombinant optical probes for cell imaging. Chembiochem. 2010 Feb 15;11(3):345-50. PubMed, PubMedCentral, CrossRef
  19. Miao Z, Ren G, Liu H, Jiang L, Cheng Z. Cy5.5-labeled Affibody molecule for near-infrared fluorescent optical imaging of epidermal growth factor receptor positive tumors. J Biomed Opt. 2010 May-Jun;15(3):036007. PubMed, CrossRef
  20. Westerlund K, Honarvar H, Tolmachev V, Eriksson Karlström A. Design, Preparation, and Characterization of PNA-Based Hybridization Probes for Affibody-Molecule-Mediated Pretargeting. Bioconjug Chem. 2015 Aug 19;26(8):1724-36. PubMed, CrossRef
  21. Hwang H, Vreven T, Weng Z. Binding interface prediction by combining protein-protein docking results. Proteins. 2014 Jan;82(1):57-66. PubMed, PubMedCentral, CrossRef
  22. Ohue M, Shimoda T, Suzuki S, Matsuzaki Y, Ishida T, Akiyama Y. MEGADOCK 4.0: an ultra-high-performance protein-protein docking software for heterogeneous supercomputers. Bioinformatics. 2014 Nov 15;30(22):3281-3. PubMed, PubMedCentral, CrossRef
  23. Wållberg H, Orlova A, Altai M, Hosseinimehr SJ, Widström C, Malmberg J, Ståhl S, Tolmachev V. Molecular design and optimization of 99mTc-labeled recombinant affibody molecules improves their biodistribution and imaging properties. J Nucl Med. 2011 Mar;52(3):461-9. PubMed, CrossRef
  24. Eigenbrot C, Ultsch M, Dubnovitsky A, Abrahmsén L, Härd T. Structural basis for high-affinity HER2 receptor binding by an engineered protein. Proc Natl Acad Sci USA. 2010 Aug 24;107(34):15039-44.  PubMed, PubMedCentralCrossRef
  25. Kelley L, Jefferys R. “Phyre2: Protein homology/analogy recognition engine V 2.0”. Structural Bioinformatics Group, Imperial College, London. 22 April 2011.
  26. Ausubel FM, Brent R, Kingston RE, Moore DD, Seilman JG, Smith JA, Struhi R. Current protocols in molecular biology (John Wiley & Sons, Inc), 1991.
  27. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680-5. PubMed, CrossRef

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