Biotechnology and Bioprocess Engineering

, Volume 20, Issue 2, pp 211–217 | Cite as

Recombinant mussel coating protein fused with cell adhesion recognition motif enhanced cell proliferation

  • Hee Young Yoo
  • Young Hoon Song
  • Jeong Hyun Seo
  • Hyung Joon Cha
  • Dong Soo Hwang
Research Paper

Abstract

Endurable coating on biomedical substrates is one of the most important issues in biomedical engineering field. Recently, a robust yet reversible coating has found in the cuticle of mussel byssus and the mimicking of the mussel cuticle coating has been considered as a strategy to build up endurable coating on biomedical materials. To date, the only known protein in the cuticle is mussel foot protein-1 (fp-1). To form endurable and bioactive coating on the biomedical surface, the fusion protein of fp-1 and GRGDSP peptide (fp-1-RGD) was genetically designed and produced in E. coli. The fusion protein of fp-1-RGD was successfully expressed as a form of inclusion body and was simply purified by diluted acetic acid extraction with high purity (~95%). Fp-1-RGD was coated on the tissue culture polystyrene (TCPS) and showed better preosteoblast cell proliferation than that of TCPS. Therefore, the marriage of fp-1 and bioactive peptide can be a good strategy to form bioactive and endurable coating in biomedical field.

Keywords

biomedical coating material mussel coating protein RGD peptide cell recognition motif fusion protein 

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References

  1. 1.
    Sun, C.and J. Waite (2005) Mapping chemical gradients within and along a fibrous structural tissue: mussel byssal threads. J. Biol. Chem. 280: 39332.CrossRefGoogle Scholar
  2. 2.
    Qin, X.and J. Waite (1998) A collagenous precursor that may mediate block copolymer gradients in mussel byssal threads. Proc. Natl. Acad. Sci. USA. 95: 10517.CrossRefGoogle Scholar
  3. 3.
    Lee, B. P., P. B. Messersmith, J. N. Israelachvili, and J. H. Waite (2011) Mussel-inspired adhesives and coatings. Annu. Rev. Mater. Res. 41: 99–132.CrossRefGoogle Scholar
  4. 4.
    Tamarin, A., P. Lewis, and J. Askey (1976) The structure and formation of the byssus attachment plaque in Mytilus. J. Morphol. 149: 199–221.CrossRefGoogle Scholar
  5. 5.
    Floriolli, R., J. von Langen, and J. Waite (2000) Marine surfaces and the expression of specific byssal adhesive protein variants in Mytilus. Mar. Biotechnol. 2: 352.Google Scholar
  6. 6.
    Waite, J., N. Holten-Andersen, S. Jewhurst, and C. Sun (2005) Mussel adhesion: finding the tricks worth mimicking. J. Adhes. 81: 297.CrossRefGoogle Scholar
  7. 7.
    Coyne, K. J., X-X. Qin, and J. H. Waite (1997) Extensible collagen in mussel byssus: a natural block copolymer. Science. 277: 1830–1832.CrossRefGoogle Scholar
  8. 8.
    Sun, C., J. M. Lucas, and J. H. Waite (2002) Collagen-binding matrix proteins from elastomeric extraorganismic byssal fibers. Biomacromolecules. 3:1240–1248.CrossRefGoogle Scholar
  9. 9.
    Sagert, J.and J. Waite (2009) Hyperunstable matrix proteins in the byssus of Mytilus galloprovincialis. J. Exp. Biol. 210: 2224.CrossRefGoogle Scholar
  10. 10.
    Zhao, H., Waite, J.(2006) Linking adhesive and structural proteins in the attachment plaque of Mytilus californianus. J. Biol. Chem. 281: 26150.CrossRefGoogle Scholar
  11. 11.
    Rzepecki, L., K. Hansen, and J. Waite (1992) Characterization of a cystine-rich polyphenolic protein family from the blue mussel Mytilus edulis L. Biol. Bull. 183: 123.CrossRefGoogle Scholar
  12. 12.
    Inoue, K., Y. Takeuchi, D. Miki, and S. Odo (1995) Mussel adhesive plaque protein gene is a novel member of epidermal growth factor-like gene family. J. Biol. Chem. 270: 6698–6701.CrossRefGoogle Scholar
  13. 13.
    Yu, J., W. Wei, E. Danner, J. N. Israelachvili, and J. H. Waite (2011) Effects of interfacial redox in mussel adhesive protein films on mica. Adv. Mater. 23: 2362–2366.CrossRefGoogle Scholar
  14. 14.
    Yu, M., J. Hwang, and T. Deming (1999) Role of L-3,4-dihydroxyphenylalanine in mussel adhesive proteins. J. Am. Chem. Soc. 121: 5825.CrossRefGoogle Scholar
  15. 15.
    Hwang, D. S., H. Zeng, A. Masic, M. J. Harrington, J. N. Israelachvili, and J. H. Waite (2010) Protein- and metal-dependent interactions of a prominent protein in mussel adhesive plaques. J. Biol. Chem. 285: 25850–25858.CrossRefGoogle Scholar
  16. 16.
    Zhao, H., J. Sagert, D. S. Hwang, and J. H. Waite (2009) Glycosylated hydroxytryptophan in a mussel adhesive protein from Perna viridis. J. Biol. Chem. 284: 23344–23352.CrossRefGoogle Scholar
  17. 17.
    Holten-Andersen, N., G. E. Fantner, S. Hohlbauch, J. H. Waite, and F. W. Zok (2007) Protective coatings on extensible biofibres. Nat. Mater. 6: 669–672.CrossRefGoogle Scholar
  18. 18.
    Holten-Andersen, N., T. E. Mates, M. S. Toprak, G. D. Stucky, F. W. Zok, and J. H. Waite (2009) Metals and the integrity of a biological coating: the cuticle of mussel byssus. Langmuir. 25: 3323–3326.CrossRefGoogle Scholar
  19. 19.
    Waite, J.(1983) Evidence for a repeated DOPA- and hydroxyproline containing decapeptide in the adhesive protein of the mussel Mytilus edulis. J. Biol. Chem. 258: 2911.Google Scholar
  20. 20.
    Taylor, S., J. Waite, M. Ross, J. Shabanowitz, and D. Hunt (1994) trans-2,3-cis-3,4-Dihydroxyproline in the tandemly repeated consensus decapeptides of an adhesive protein from Mytilus edulis. J. Am. Chem. Soc. 116: 10803.CrossRefGoogle Scholar
  21. 21.
    Filpula, D., S. Lee, R. Link, S. Strausberg, and R. Strausberg (1990) Structural and functional repetition in a marine mussel adhesive protein. Biotechnol. Prog. 6: 171.CrossRefGoogle Scholar
  22. 22.
    Hwang, D. S. and J. H. Waite (2012) Three intrinsically unstructured mussel adhesive proteins, mfp-1, mfp-2, and mfp-3: Analysis by circular dichroism. Protein Sci. 21: 1689–1695.CrossRefGoogle Scholar
  23. 23.
    Kanyalkar, M., S. Srivastava, and E. Coutinho (2002) Conformation of a model peptide of the tandem repeat decapeptide in mussel adhesive protein by NMR and MD simulations. Biomaterials. 23: 389–396.CrossRefGoogle Scholar
  24. 24.
    Anderson, K.and J. Waite (2000) Immunolocalization of Dpfp1, a byssal protein of the zebra mussel (Dreissena polymorpha). J. Exp. Biol. 203: 3065.Google Scholar
  25. 25.
    Zeng, H., D. S. Hwang, J. N. Israelachvili, and J. H. Waite (2010) Strong reversible Fe3+-mediated bridging between dopa-containing protein films in water. Proc. Natl. Acad. Sci. USA. 107: 12850–12853.CrossRefGoogle Scholar
  26. 26.
    Hynes, R.O. (2002) Integrins: bidirectional, allosteric signaling machines. Cell. 110: 673–687.CrossRefGoogle Scholar
  27. 27.
    Frisch, S.and H. Francis (1994) Disruption of epithelial cellmatrix interactions induces apoptosis. J. Cell Biol. 124: 619–626.CrossRefGoogle Scholar
  28. 28.
    Giancotti, F. G. and E. Ruoslahti (1999) Integrin signaling. Science. 285: 1028–1033.CrossRefGoogle Scholar
  29. 29.
    Chua, P-H., K-G. Neoh, E-T. Kang, and W. Wang (2008) Surface functionalization of titanium with hyaluronic acid/chitosan polyelectrolyte multilayers and RGD for promoting osteoblast functions and inhibiting bacterial adhesion. Biomaterials. 29: 1412–1421.CrossRefGoogle Scholar
  30. 30.
    Ruoslahti, E.(1996) RGD and other recognition sequences for integrins. Ann. Rev. Cell Dev. Biol. 12: 697–715.Google Scholar
  31. 31.
    Hwang, D. S., S. B. Sim, and H. J. Cha. (2007) Cell adhesion biomaterial based on mussel adhesive protein fused with RGD peptide. Biomaterials. 28: 4039–4046.CrossRefGoogle Scholar
  32. 32.
    Rzepecki, L. M., S. S. Chin, J. H. Waite, and M. F. Lavin (1991) Molecular diversity of marine glues: polyphenolic proteins from five mussel species. Mol. Mar. Biol. Biotechnol. 1: 78–88.Google Scholar
  33. 33.
    Hwang, D. S., H. J. Yoo, J. H. Jun, W. K. Moon, and H. J. Cha (2004) Expression of functional recombinant mussel adhesive protein Mgfp-5 in Escherichia coli. Appl. Environ. Microbiol. 70: 3352–3359.CrossRefGoogle Scholar
  34. 34.
    Hwang, D. S., Y. Gim, and H. J. Cha (2005) Expression of functional recombinant mussel adhesive protein type 3A in Escherichia coli. Biotechnol. Prog. 21: 965–970.CrossRefGoogle Scholar
  35. 35.
    Waite, J.H. (1995) Precursors of quinone tanning: dopa-containing proteins. Methods Enzymol. 258: 1–20.CrossRefGoogle Scholar
  36. 36.
    Magalhaes, P. O., A. M. Lopes, P. G. Mazzola, C. Rangel-Yagui, T. C. V. Penna, and A. Pessoa (2007) Methods of endotoxin removal from biological preparations: a review. J. Pharm. Pharm. Sci. 10: 388–404.Google Scholar
  37. 37.
    Papov, V. V., T. V. Diamond, K. Biemann, and J. H. Waite (1995) Hydroxyarginine-containing polyphenolic proteins in the adhesive plaques of the marine mussel Mytilus edulis. J. Biol. Chem. 270: 20183–20192.CrossRefGoogle Scholar
  38. 38.
    Kamino, K., K. Inoue, T. Maruyama, N. Takamatsu, S. Harayama, and Y. Shizuri (2000) Barnacle cement proteins: importance of disulfide bonds in their insolubility. J. Biol. Chem.Google Scholar
  39. 39.
    Tominaga, H., M. Ishiyama, F. Ohseto, K. Sasamoto, T. Hamamoto, K. Suzuki, and M. Watanabe (1999) A water-soluble tetrazolium salt useful for colorimetric cell viability assay. Anal. Comm. 36: 47–50.CrossRefGoogle Scholar
  40. 40.
    Ruoslahti, E.and M. Pierschbacher (1987) New perspectives in cell adhesion: RGD and integrins. Science. 238: 491–497.CrossRefGoogle Scholar
  41. 41.
    Dalgleish, R.(1997) The human type I collagen mutation database. Nucleic Acids Res. 25: 181–187.Google Scholar
  42. 42.
    Rossert, J., C. Terraz, and S. Dupont (2000) Regulation of type I collagen genes expression. Nephrol. Dial. Transplant. 15(suppl 6): 66–68.CrossRefGoogle Scholar
  43. 43.
    Kim, J-B., P. Leucht, C. A. Luppen, Y. J. Park, H. E. Beggs, C. H. Damsky, and J. A. Helms (2007) Reconciling the roles of FAK in osteoblast differentiation, osteoclast remodeling, and bone regeneration. Bone. 41: 39–51.CrossRefGoogle Scholar
  44. 44.
    Heino, J.(2000) The collagen receptor integrins have distinct ligand recognition and signaling functions. Matrix Biol. 19: 319–323.Google Scholar

Copyright information

© The Korean Society for Biotechnology and Bioengineering and Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Hee Young Yoo
    • 1
  • Young Hoon Song
    • 2
  • Jeong Hyun Seo
    • 2
  • Hyung Joon Cha
    • 3
  • Dong Soo Hwang
    • 1
    • 4
  1. 1.Integrative Bioscience and BioengineeringPohang University of Science and TechnologyPohangKorea
  2. 2.School of Chemical EngineeringYeungnam UniversityGyeongsanKorea
  3. 3.Department of Chemical EngineeringPohang University of Science and TechnologyPohangKorea
  4. 4.School of Environmental Science and EngineeringPohang University of Science and TechnologyPohangKorea

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