Spandidos Publications Logo
  • About
    • About Spandidos
    • Aims and Scopes
    • Abstracting and Indexing
    • Editorial Policies
    • Reprints and Permissions
    • Job Opportunities
    • Terms and Conditions
    • Contact
  • Journals
    • All Journals
    • Oncology Letters
      • Oncology Letters
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Oncology
      • International Journal of Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular and Clinical Oncology
      • Molecular and Clinical Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Experimental and Therapeutic Medicine
      • Experimental and Therapeutic Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Molecular Medicine
      • International Journal of Molecular Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Biomedical Reports
      • Biomedical Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Reports
      • Oncology Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular Medicine Reports
      • Molecular Medicine Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • World Academy of Sciences Journal
      • World Academy of Sciences Journal
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Functional Nutrition
      • International Journal of Functional Nutrition
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Epigenetics
      • International Journal of Epigenetics
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Medicine International
      • Medicine International
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
  • Articles
  • Information
    • Information for Authors
    • Information for Reviewers
    • Information for Librarians
    • Information for Advertisers
    • Conferences
  • Language Editing
Spandidos Publications Logo
  • About
    • About Spandidos
    • Aims and Scopes
    • Abstracting and Indexing
    • Editorial Policies
    • Reprints and Permissions
    • Job Opportunities
    • Terms and Conditions
    • Contact
  • Journals
    • All Journals
    • Biomedical Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Experimental and Therapeutic Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Epigenetics
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Functional Nutrition
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Molecular Medicine
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • International Journal of Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Medicine International
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular and Clinical Oncology
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Molecular Medicine Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Letters
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • Oncology Reports
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
    • World Academy of Sciences Journal
      • Information for Authors
      • Editorial Policies
      • Editorial Board
      • Aims and Scope
      • Abstracting and Indexing
      • Bibliographic Information
      • Archive
  • Articles
  • Information
    • For Authors
    • For Reviewers
    • For Librarians
    • For Advertisers
    • Conferences
  • Language Editing
Login Register Submit
  • This site uses cookies
  • You can change your cookie settings at any time by following the instructions in our Cookie Policy. To find out more, you may read our Privacy Policy.

    I agree
Search articles by DOI, keyword, author or affiliation
Search
Advanced Search
presentation
Molecular Medicine Reports
Join Editorial Board Propose a Special Issue
Print ISSN: 1791-2997 Online ISSN: 1791-3004
Journal Cover
April-2016 Volume 13 Issue 4

Full Size Image

Sign up for eToc alerts
Recommend to Library

Journals

International Journal of Molecular Medicine

International Journal of Molecular Medicine

International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

International Journal of Oncology

International Journal of Oncology

International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

Molecular Medicine Reports

Molecular Medicine Reports

Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery.

Oncology Reports

Oncology Reports

Oncology Reports is an international journal devoted to fundamental and applied research in Oncology.

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

Oncology Letters

Oncology Letters

Oncology Letters is an international journal devoted to Experimental and Clinical Oncology.

Biomedical Reports

Biomedical Reports

Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology.

Molecular and Clinical Oncology

Molecular and Clinical Oncology

International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis.

World Academy of Sciences Journal

World Academy of Sciences Journal

Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology.

International Journal of Functional Nutrition

International Journal of Functional Nutrition

Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition.

International Journal of Epigenetics

International Journal of Epigenetics

Publishes open-access research on using epigenetics to advance understanding and treatment of human disease.

Medicine International

Medicine International

An International Open Access Journal Devoted to General Medicine.

Journal Cover
April-2016 Volume 13 Issue 4

Full Size Image

Sign up for eToc alerts
Recommend to Library

  • Article
  • Citations
    • Cite This Article
    • Download Citation
    • Create Citation Alert
    • Remove Citation Alert
    • Cited By
  • Similar Articles
    • Related Articles (in Spandidos Publications)
    • Similar Articles (Google Scholar)
    • Similar Articles (PubMed)
  • Download PDF
  • Download XML
  • View XML
Article

Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration

  • Authors:
    • Yonghuan Song
    • Jian Ding
    • Rilong Jin
    • Jinhee Jung
    • Shi Li
    • Jingquan Yang
    • Anyuan Wang
    • Zhijie Li
  • View Affiliations / Copyright

    Affiliations: Department of Hand and Plastic Surgery, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, P.R. China, Department of Orthopedic Surgery, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310000, P.R. China, Molecular Evolution Team, Department of Biotech R&D, Amicogen Inc., Jinju, Yeongnam 660‑852, South Korea
  • Pages: 3619-3626
    |
    Published online on: February 26, 2016
       https://doi.org/10.3892/mmr.2016.4942
  • Expand metrics +
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )
Cited By (CrossRef): 0 citations Loading Articles...

This article is mentioned in:



Abstract

Fibroblast growth factor (FGF)21 functions in the maintenance of glucose homeostasis and exerts protective effects on the liver, heat and kidneys. However, the roles of FGF21 in other tissue types are yet to be fully elucidated. The present study detected elevated expression levels of FGF21 in skin tissue. Furthermore, it was revealed that FGF21 expression in the skin was induced upon wounding. In addition, β‑klotho expression was detected in the skin tissue. To examine the role of FGF21 in the wound healing process, recombinant human (h)FGF21 was expressed in a the yeast strain Pichia (P.) pastoris, a well‑known system for recombinant protein production. Based on the sequence of hFGF21 and the optimal codon of P. pastoris, codon‑optimized FGF21 open reading frame sequences were obtained using seven pairs of 55‑59‑nt primers with seven rounds of PCR. The recombinant FGF21 was purified and its function was examined in human fibroblast cells using a wound healing cell migration assay. Treatment with FGF21 promoted cell migration, which is an important step in wound healing. Furthermore, FGF21 treatment enhanced the activity of c‑Jun N‑terminal kinase, a key regulator in fibroblast‑cell migration. In conclusion, FGF21 is induced after wounding and FGF21 expressed and purified from yeast markedly accelerates wound healing. The present study was the first to elucidate the function of FGF21 in skin tissues and provided a theoretical basis for the use of FGF21 in the treatment of skin wounds.
View Figures

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

View References

1 

Mohammadi M, Olsen SK and Ibrahimi OA: Structural basis for fibroblast growth factor receptor activation. Cytokine Growth Factor Rev. 16:107–137. 2005. View Article : Google Scholar : PubMed/NCBI

2 

Feldman B, Poueymirou W, Papaioannou VE, DeChiara TM and Goldfarb M: Requirement of FGF-4 for postimplantation mouse development. Science. 267:246–249. 1995. View Article : Google Scholar : PubMed/NCBI

3 

Dubrulle J and Pourquié O: Fgf8 mRNA decay establishes a gradient that couples axial elongation to patterning in the vertebrate embryo. Nature. 427:419–422. 2004. View Article : Google Scholar : PubMed/NCBI

4 

Sun X, Meyers EN, Lewandoski M and Martin GR: Targeted disruption of Fgf8 causes failure of cell migration in the gastrulating mouse embryo. Genes Dev. 13:1834–1846. 1999. View Article : Google Scholar : PubMed/NCBI

5 

Martin GR: The roles of FGFs in the early development of vertebrate limbs. Genes Dev. 12:1571–1586. 1998. View Article : Google Scholar : PubMed/NCBI

6 

Goldfarb M: Functions of fibroblast growth factors in vertebrate development. Cytokine Growth Factor Rev. 7:311–325. 1996. View Article : Google Scholar : PubMed/NCBI

7 

Kanazawa S, Fujiwara T, Matsuzaki S, Shingaki K, Taniguchi M, Miyata S, Tohyama M, Sakai Y, Yano K, Hosokawa K and Kubo T: BFGF regulates PI3-kinase-Rac1-JNK pathway and promotes fibroblast migration in wound healing. PLoS One. 5:e122282010. View Article : Google Scholar : PubMed/NCBI

8 

Nishimura T, Nakatake Y, Konishi M and Itoh N: Identification of a novel FGF, FGF-21, preferentially expressed in the liver. Biochim Biophys Acta. 1492:203–206. 2000. View Article : Google Scholar : PubMed/NCBI

9 

Liang Q, Zhong L, Zhang J, Wang Y, Bornstein SR, Triggle CR, Ding H, Lam KS and Xu A: FGF21 maintains glucose homeostasis by mediating the cross talk between liver and brain during prolonged fasting. Diabetes. 63:4064–4075. 2014. View Article : Google Scholar : PubMed/NCBI

10 

Lin Z, Tian H, Lam KS, Lin S, Hoo RC, Konishi M, Itoh N, Wang Y, Bornstein SR, Xu A and Li X: Adiponectin mediates the metabolic effects of FGF21 on glucose homeostasis and insulin sensitivity in mice. Cell Metab. 17:779–789. 2013. View Article : Google Scholar : PubMed/NCBI

11 

Lin Z, Wu F, Lin S, Pan X, Jin L, Lu T, Shi L, Wang Y, Xu A and Li X: Adiponectin protects against acetaminophen-induced mitochondrial dysfunction and acute liver injury by promoting autophagy in mice. J Hepatol. 61:825–831. 2014. View Article : Google Scholar : PubMed/NCBI

12 

Yie J, Wang W, Deng L, Tam LT, Stevens J, Chen MM, Li Y, Xu J, Lindberg R, Hecht R, et al: Understanding the physical interactions in the FGF21/FGFR/β-Klotho complex: Structural requirements and implications in FGF21 signaling. Chem Biol Drug Des. 79:398–410. 2012. View Article : Google Scholar : PubMed/NCBI

13 

Belov AA and Mohammadi M: Molecular mechanisms of fibroblast growth factor signaling in physiology and pathology. Cold Spring Harb Perspect Biol. 5:a0159582013. View Article : Google Scholar : PubMed/NCBI

14 

Wang H, Xiao Y, Fu L, Zhao H, Zhang Y, Wan X, Qin Y, Huang Y, Gao H and Li X: High-level expression and purification of soluble recombinant FGF21 protein by SUMO fusion in Escherichia coli. BMC Biotechnol. 10:142010. View Article : Google Scholar : PubMed/NCBI

15 

Daly R and Hearn MT: Expression of heterologous proteins in Pichia pastoris: A useful experimental tool in protein engineering and production. J Mol Recognit. 18:119–138. 2005. View Article : Google Scholar

16 

Zittermann SI and Issekutz AC: Basic fibroblast growth factor (bFGF, FGF-2) potentiates leukocyte recruitment to inflammation by enhancing endothelial adhesion molecule expression. Am J Pathol. 168:835–846. 2006. View Article : Google Scholar : PubMed/NCBI

17 

Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods. 25:402–408. 2001. View Article : Google Scholar

18 

Jin F, Xu X, Zhang W and Gu D: Expression and characterization of a housefly cecropin gene in the methylotrophic yeast, Pichia pastoris. Protein Expr Purif. 49:39–46. 2006. View Article : Google Scholar : PubMed/NCBI

19 

Jin FL, Xu XX, Yu XQ and Ren SX: High-level expression of active recombinant ubiquitin carboxyl-terminal hydrolase of Drosophila melanogaster in Pichia pastoris. Protein Expr Purif. 65:115–121. 2009. View Article : Google Scholar

20 

Xuan YH, Huang BB, Tian HS, Chi LS, Duan YM, Wang X, Zhu ZX, Cai WH, Zhu YT, Wei TM, et al: High-glucose inhibits human fibroblast cell migration in wound healing via repression of bFGF-regulating JNK phosphorylation. PLoS One. 9:e1081822014. View Article : Google Scholar : PubMed/NCBI

21 

Iyer VR, Eisen MB, Ross DT, Schuler G, Moore T, Lee JC, Trent JM, Staudt LM, Hudson J Jr, Boguski MS, et al: The transcriptional program in the response of human fibroblasts to serum. Science. 283:83–87. 1999. View Article : Google Scholar : PubMed/NCBI

22 

Burgess WH and Maciag T: The heparin-binding (fibroblast) growth factor family of proteins. Annu Rev Biochemistry. 58:575–602. 1989. View Article : Google Scholar

23 

Courty J, Dauchel MC, Caruelle D, Perderiset M and Barritault D: Mitogenic properties of a new endothelial cell growth factor related to pleiotrophin. Biochem Biophys Res Commun. 180:145–151. 1991. View Article : Google Scholar : PubMed/NCBI

24 

Fernig DG and Gallagher JT: Fibroblast growth factors and their receptors: An information network controlling tissue growth, morphogenesis and repair. Prog Growth Factor Res. 5:353–377. 1994. View Article : Google Scholar : PubMed/NCBI

25 

Huang Z, Wang H, Lu M, Sun C, Wu X, Tan Y, Ye C, Zhu G, Wang X, Cai L and Li X: A better anti-diabetic recombinant human fibroblast growth factor 21 (rhFGF21) modified with polyethylene glycol. PLoS One. 6:e206692011. View Article : Google Scholar : PubMed/NCBI

26 

Kharitonenkov A, Shiyanova TL, Koester A, Ford AM, Micanovic R, Galbreath EJ, Sandusky GE, Hammond LJ, Moyers JS, Owens RA, et al: FGF-21 as a novel metabolic regulator. J Clin Invest. 115:1627–1635. 2005. View Article : Google Scholar : PubMed/NCBI

27 

Gu W, Zhou T and Wilke CO: A universal trend of reduced mRNA stability near the translation-initiation site in prokaryotes and eukaryotes. PLoS Comput Biol. 6:e1000664. 2010. View Article : Google Scholar : PubMed/NCBI

28 

Zhang M, Jiang X, Su Z, Lin J, Xiang Q, Yang Z, Huang Y and Li X: Large-scale expression, purification, and glucose uptake activity of recombinant human FGF21 in Escherichia coli. Appl Microbiol Biotechnol. 93:613–621. 2012. View Article : Google Scholar

29 

Martin P: Wound healing-aiming for perfect skin regeneration. Science. 276:75–81. 1997. View Article : Google Scholar : PubMed/NCBI

30 

Cao C, Sun Y, Healey S, Bi Z, Hu G, Wan S, Kouttab N, Chu W and Wan Y: EGFR-mediated expression of aquaporin-3 is involved in human skin fibroblast migration. Biochem J. 400:225–234. 2006. View Article : Google Scholar : PubMed/NCBI

31 

Dever TE: Using GCN4 as a reporter of eIF2 alpha phosphorylation and translational regulation in yeast. Methods. 11:403–417. 1997. View Article : Google Scholar : PubMed/NCBI

32 

Payne T, Finnis C, Evans LR, Mead DJ, Avery SV, Archer DB and Sleep D: Modulation of chaperone gene expression in mutagenized Saccharomyces cerevisiae strains developed for recombinant human albumin production results in increased production of multiple heterologous proteins. Appl Environ Microbiol. 74:7759–7766. 2008. View Article : Google Scholar : PubMed/NCBI

33 

Kazemi Seresht A, Nørgaard P, Palmqvist EA, Andersen AS and Olsson L: Modulating heterologous protein production in yeast: The applicability of truncated auxotrophic markers. Appl Microbiol Biotechnol. 97:3939–3948. 2013. View Article : Google Scholar

Related Articles

  • Abstract
  • View
  • Download
  • Twitter
Copy and paste a formatted citation
Spandidos Publications style
Song Y, Ding J, Jin R, Jung J, Li S, Yang J, Wang A and Li Z: Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration. Mol Med Rep 13: 3619-3626, 2016.
APA
Song, Y., Ding, J., Jin, R., Jung, J., Li, S., Yang, J. ... Li, Z. (2016). Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration. Molecular Medicine Reports, 13, 3619-3626. https://doi.org/10.3892/mmr.2016.4942
MLA
Song, Y., Ding, J., Jin, R., Jung, J., Li, S., Yang, J., Wang, A., Li, Z."Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration". Molecular Medicine Reports 13.4 (2016): 3619-3626.
Chicago
Song, Y., Ding, J., Jin, R., Jung, J., Li, S., Yang, J., Wang, A., Li, Z."Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration". Molecular Medicine Reports 13, no. 4 (2016): 3619-3626. https://doi.org/10.3892/mmr.2016.4942
Copy and paste a formatted citation
x
Spandidos Publications style
Song Y, Ding J, Jin R, Jung J, Li S, Yang J, Wang A and Li Z: Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration. Mol Med Rep 13: 3619-3626, 2016.
APA
Song, Y., Ding, J., Jin, R., Jung, J., Li, S., Yang, J. ... Li, Z. (2016). Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration. Molecular Medicine Reports, 13, 3619-3626. https://doi.org/10.3892/mmr.2016.4942
MLA
Song, Y., Ding, J., Jin, R., Jung, J., Li, S., Yang, J., Wang, A., Li, Z."Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration". Molecular Medicine Reports 13.4 (2016): 3619-3626.
Chicago
Song, Y., Ding, J., Jin, R., Jung, J., Li, S., Yang, J., Wang, A., Li, Z."Expression and purification of FGF21 in Pichia pastoris and its effect on fibroblast-cell migration". Molecular Medicine Reports 13, no. 4 (2016): 3619-3626. https://doi.org/10.3892/mmr.2016.4942
Follow us
  • Twitter
  • LinkedIn
  • Facebook
About
  • Spandidos Publications
  • Careers
  • Cookie Policy
  • Privacy Policy
How can we help?
  • Help
  • Live Chat
  • Contact
  • Email to our Support Team