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Research article

References

  1. Carpenter G, Cohen S. 1976. 125I-labeled human epidermal growth factor. Binding internalization, and degradation in human fibroblast. J. Cell Biol. 71: 159-171.
    Pubmed CrossRef
  2. Carpenter G, Cohen S. 1979. Epidermal growth factor. Ann. Rev. Biochem. 48: 193-216.
    Pubmed CrossRef
  3. Chen X, Cen P, Chen J. 2005. Enhanced production of human epidermal growth factor by a recombinant Escherichia coli integrated with in situ exchange of acetic acid by macroporous ion-exchange resin. J. Biosci. Bioeng. 5: 579-581.
    Pubmed CrossRef
  4. Chung KH, Park SH, Kim MK, Park HD, Son TI. 2006. Stabilization of epidermal growth factor on thermal and proteolytic degradation by conjugation with low molecular weight chitosan. J. Appl. Polym. Sci. 102: 5072-5082.
    CrossRef
  5. Dagmar P, Anspach FB. 2000. Endotoxin removal from protein solution. J. Biotechnol. 76: 97-119.
    CrossRef
  6. Egawa EY, Kato K, Hiraoka M, Nakaji-Hirabayashi T, Iwata H. 2011. Enhanced proliferation of neural stem cells in a collagen hydrogel incorporating engineered epidermal growth factor. Biomaterials 32: 4737-4743.
    Pubmed CrossRef
  7. Elloumi I, Kobayashi R, Funabashi H, Mie M, Kobatake E. 2006. Construction of epidermal growth factor fusion protein with cell adhesive activity. Biomaterials 27: 3451-3458.
    Pubmed CrossRef
  8. Elloumi HI, Nakamura M, Mie M, Kobatake E. 2009. Construction of multifunctional proteins for tissue engineering:epidermal growth factor with collagen binding and cell adhesive activities. J. Biotechnol. 139: 19-25.
    Pubmed CrossRef
  9. Hardwicke J, Schmaljohann D, Boyce D, Thomas D. 2008. Epidermal growth factor therapy and wound healing – past, present and future perspective. Surgeon 6: 172-177.
    CrossRef
  10. Hayashi M, Tomita M, Yoshizato K. 2001. Production of EGF-collagen chimeric protein which shows the mitogenic activity. Biochem. Biophys. Acta 1528: 187-195.
    CrossRef
  11. Imanishi J, Kamiyama K, Iguchi I, Kita M, Sotozono C, Kinoshita S. 2000. Growth factors: importance in wound healing and maintenance of transparence of the cornea. Prog. Retin. Eye Res. 19: 113-129.
    CrossRef
  12. Ishikawa T, Terai H, Kitajima T. 2001. Production of a biologically active epidermal growth factor fusion protein with high collagen affinity. J. Biochem. 129: 627-633.
    Pubmed CrossRef
  13. Ishikawa T, Terai H, Yamamoto T, Harada K, Kitajima T. 2003. Delivery of a growth factor fusion protein having collagen-binding activity to wound tissues. Artif. Organs 27:147-154.
    Pubmed CrossRef
  14. Kim DG, Min MK, Ahn SC, Kim JK, Kong IS. 2009. Expression of a fusion protein containing human epidermal growth factor and the collagen-binding domain of Vibrio mimicus. Biotechnol. Lett. 31: 259-264.
    Pubmed CrossRef
  15. Kitajima T, Makoto S, Hirokazu H, Takeshi O, Yoshihiro I. 2009. A chimeric epidermal growth factor with fibrin affinity promotes repair of injured keratinocyte sheets. Acta Biomater. 5: 2623-2632.
    Pubmed CrossRef
  16. Lee JH, Ahn SH, Lee EM, Jeong SH, Kim YO, Lee SJ, Kong IS. 2005. The FAXWXXT motif in the carboxyl terminus of Vibrio mimicus metalloprotease is involved in binding to collagen. FEBS Lett. 579: 2507-2513.
    Pubmed CrossRef
  17. Liu YL, Huang LM, Lin WP, Tsai CC, Lin TS, Hu YH, et al. 2006. Secretion of biologically active human epidermal growth factor from Escherichia coli using Yersinia pestis Caf1 signal peptide. J. Microbiol. Immunol. Infect. 39: 366-371.
    Pubmed
  18. Nakagawa S, Yoshida S, Hirao Y, Kasuga S, Fuwa T. 1985. Biological effects of biosynthetic human EGF on the growth of mammalian cell in vitro. Differentiation 29: 284-288.
    Pubmed CrossRef
  19. Nimni ME. 1997. Polypeptide growth factors: targeted delivery systems. Biomaterials 18: 1201-1225.
    CrossRef
  20. Nishi N, Matsushita O, Yuube K, Miyanaka H, Okabe A, Wada F. 1998. Collagen-binding growth factors: production and characterization of functional fusion protein having a collagen-binding domain. Proc. Natl. Acad. Sci. USA 95: 70187023.
    CrossRef
  21. Rustad KC, Sorkin M, Levi B, Longaker MT, Gurtner GC. 2010. Strategies for organ level tissue engineering. Organogenesis 6: 151-157.
    Pubmed PMC CrossRef
  22. Sahasrabuddhe AA, Ahmed N, Krishnasastry MV. 2006. Stress-induced phosphorylation of caveolin-1 and p38, and down-regulation of EGFr and ERK by the dietary lectin jacalin in two human carcinoma cell lines. Cell Stress Chaperones 11: 135-147.
    Pubmed PMC CrossRef
  23. Schlessinger J, Schreiber AB, Levi A, Lax I, Libermann T, Yarden Y. 1983. Regulation of cell proliferation by epidermal growth factor. CRC Crit. Rev. Biochem. 14: 93-111.
    Pubmed CrossRef
  24. Yang Y, Zhao Y, Chen B, Han Q, Sun W, Xio Z, Dai J. 2009. Collagen-binding human epidermal growth factor promotes cellularization of collagen scaffolds. Tissue Eng. A 15: 35893596.
    Pubmed CrossRef
  25. Yin J, Xu K, Zhang J, Kumar A, Yu FX. 2007. Woundinduced ATP release and EGF receptor activation in epidermal cells. J. Cell Sci. 120: 815-825.
    Pubmed PMC CrossRef

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Article

Research article

J. Microbiol. Biotechnol. 2015; 25(1): 119-126

Published online January 28, 2015 https://doi.org/10.4014/jmb.1405.05073

Copyright © The Korean Society for Microbiology and Biotechnology.

Construction of Chimeric Human Epidermal Growth Factor Containing Short Collagen-Binding Domain Moieties for Use as a Wound Tissue Healing Agent

Dong-Gyun Kim 1, Eun-Young Kim 2, Yu-Ri Kim 3 and In-Soo Kong 4*

1Biotechnology Research Division, National Fisheries Research and Development Institute, Busan 619-705, Republic of Korea, 2Institute of Fisheries Science, Pukyong National University, Busan 608-737, Republic of Korea , 3CBR Defense Research Institute, The Armed Forces CBD Defense Command, Seoul 137-180, Republic of Korea, 4Department of Biotechnology, Pukyong National University, Busan 608-737, Republic of Korea

Received: May 30, 2014; Accepted: August 18, 2014

Abstract

Among the various human growth factors, epidermal growth factor (hEGF, consisting of 53
amino acids) has various effects on cell regeneration, stimulation of proliferation, migration of
keratinocytes, formation of granulation tissues, and stimulation of fibroblast motility, which
are important for wound healing. Owing to their multiple activities, EGFs are used as
pharmaceutical and cosmetic agents. However, their low productivity, limited target
specificity, and short half-life inhibit their application as therapeutic agents. To overcome
these obstacles, we fused the collagen-binding domain (CBD) of Vibrio mimicus metalloprotease
to EGF protein. About 18 or 12 amino acids (aa) (of the 33 total amino acids), which were
essential for collagen-binding activity, were combined with the N- and C-termini of EGF. We
constructed, expressed, and purified EGF (53 aa)-CBD (18 aa), EGF (53 aa)-CBD (12 aa), CBD
(18 aa)-EGF (53 aa), and CBD (12 aa)-EGF (53 aa). These purified recombinant proteins
increased the numbers of cells in treated specimens compared with non-treated specimens and
control hEGF samples. The collagen-binding activities were also evaluated. Furthermore,
CBD-hybridized hEGF induced phosphorylation of the EGF receptor. These results suggested
that these fusion proteins could be applicable as small therapeutic agents in wound tissue
healing.

Keywords: Collagen-binding domain (CBD), fusion protein, Human epidermal growth factor (EGF), wound healing agent

References

  1. Carpenter G, Cohen S. 1976. 125I-labeled human epidermal growth factor. Binding internalization, and degradation in human fibroblast. J. Cell Biol. 71: 159-171.
    Pubmed CrossRef
  2. Carpenter G, Cohen S. 1979. Epidermal growth factor. Ann. Rev. Biochem. 48: 193-216.
    Pubmed CrossRef
  3. Chen X, Cen P, Chen J. 2005. Enhanced production of human epidermal growth factor by a recombinant Escherichia coli integrated with in situ exchange of acetic acid by macroporous ion-exchange resin. J. Biosci. Bioeng. 5: 579-581.
    Pubmed CrossRef
  4. Chung KH, Park SH, Kim MK, Park HD, Son TI. 2006. Stabilization of epidermal growth factor on thermal and proteolytic degradation by conjugation with low molecular weight chitosan. J. Appl. Polym. Sci. 102: 5072-5082.
    CrossRef
  5. Dagmar P, Anspach FB. 2000. Endotoxin removal from protein solution. J. Biotechnol. 76: 97-119.
    CrossRef
  6. Egawa EY, Kato K, Hiraoka M, Nakaji-Hirabayashi T, Iwata H. 2011. Enhanced proliferation of neural stem cells in a collagen hydrogel incorporating engineered epidermal growth factor. Biomaterials 32: 4737-4743.
    Pubmed CrossRef
  7. Elloumi I, Kobayashi R, Funabashi H, Mie M, Kobatake E. 2006. Construction of epidermal growth factor fusion protein with cell adhesive activity. Biomaterials 27: 3451-3458.
    Pubmed CrossRef
  8. Elloumi HI, Nakamura M, Mie M, Kobatake E. 2009. Construction of multifunctional proteins for tissue engineering:epidermal growth factor with collagen binding and cell adhesive activities. J. Biotechnol. 139: 19-25.
    Pubmed CrossRef
  9. Hardwicke J, Schmaljohann D, Boyce D, Thomas D. 2008. Epidermal growth factor therapy and wound healing – past, present and future perspective. Surgeon 6: 172-177.
    CrossRef
  10. Hayashi M, Tomita M, Yoshizato K. 2001. Production of EGF-collagen chimeric protein which shows the mitogenic activity. Biochem. Biophys. Acta 1528: 187-195.
    CrossRef
  11. Imanishi J, Kamiyama K, Iguchi I, Kita M, Sotozono C, Kinoshita S. 2000. Growth factors: importance in wound healing and maintenance of transparence of the cornea. Prog. Retin. Eye Res. 19: 113-129.
    CrossRef
  12. Ishikawa T, Terai H, Kitajima T. 2001. Production of a biologically active epidermal growth factor fusion protein with high collagen affinity. J. Biochem. 129: 627-633.
    Pubmed CrossRef
  13. Ishikawa T, Terai H, Yamamoto T, Harada K, Kitajima T. 2003. Delivery of a growth factor fusion protein having collagen-binding activity to wound tissues. Artif. Organs 27:147-154.
    Pubmed CrossRef
  14. Kim DG, Min MK, Ahn SC, Kim JK, Kong IS. 2009. Expression of a fusion protein containing human epidermal growth factor and the collagen-binding domain of Vibrio mimicus. Biotechnol. Lett. 31: 259-264.
    Pubmed CrossRef
  15. Kitajima T, Makoto S, Hirokazu H, Takeshi O, Yoshihiro I. 2009. A chimeric epidermal growth factor with fibrin affinity promotes repair of injured keratinocyte sheets. Acta Biomater. 5: 2623-2632.
    Pubmed CrossRef
  16. Lee JH, Ahn SH, Lee EM, Jeong SH, Kim YO, Lee SJ, Kong IS. 2005. The FAXWXXT motif in the carboxyl terminus of Vibrio mimicus metalloprotease is involved in binding to collagen. FEBS Lett. 579: 2507-2513.
    Pubmed CrossRef
  17. Liu YL, Huang LM, Lin WP, Tsai CC, Lin TS, Hu YH, et al. 2006. Secretion of biologically active human epidermal growth factor from Escherichia coli using Yersinia pestis Caf1 signal peptide. J. Microbiol. Immunol. Infect. 39: 366-371.
    Pubmed
  18. Nakagawa S, Yoshida S, Hirao Y, Kasuga S, Fuwa T. 1985. Biological effects of biosynthetic human EGF on the growth of mammalian cell in vitro. Differentiation 29: 284-288.
    Pubmed CrossRef
  19. Nimni ME. 1997. Polypeptide growth factors: targeted delivery systems. Biomaterials 18: 1201-1225.
    CrossRef
  20. Nishi N, Matsushita O, Yuube K, Miyanaka H, Okabe A, Wada F. 1998. Collagen-binding growth factors: production and characterization of functional fusion protein having a collagen-binding domain. Proc. Natl. Acad. Sci. USA 95: 70187023.
    CrossRef
  21. Rustad KC, Sorkin M, Levi B, Longaker MT, Gurtner GC. 2010. Strategies for organ level tissue engineering. Organogenesis 6: 151-157.
    Pubmed KoreaMed CrossRef
  22. Sahasrabuddhe AA, Ahmed N, Krishnasastry MV. 2006. Stress-induced phosphorylation of caveolin-1 and p38, and down-regulation of EGFr and ERK by the dietary lectin jacalin in two human carcinoma cell lines. Cell Stress Chaperones 11: 135-147.
    Pubmed KoreaMed CrossRef
  23. Schlessinger J, Schreiber AB, Levi A, Lax I, Libermann T, Yarden Y. 1983. Regulation of cell proliferation by epidermal growth factor. CRC Crit. Rev. Biochem. 14: 93-111.
    Pubmed CrossRef
  24. Yang Y, Zhao Y, Chen B, Han Q, Sun W, Xio Z, Dai J. 2009. Collagen-binding human epidermal growth factor promotes cellularization of collagen scaffolds. Tissue Eng. A 15: 35893596.
    Pubmed CrossRef
  25. Yin J, Xu K, Zhang J, Kumar A, Yu FX. 2007. Woundinduced ATP release and EGF receptor activation in epidermal cells. J. Cell Sci. 120: 815-825.
    Pubmed KoreaMed CrossRef