High production in E. coli of biologically active recombinant human fibroblast growth factor 20 and its neuroprotective effects

详细信息    查看全文

• 作者：Haishan Tian ; Yang Zhao ; Nazi Chen ; Meiyu Wu…
• 关键词：rhFGF20 ; Inclusion body protein ; Purification ; Mitogenic activity ; PC ; 12 cells
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2016
• 出版时间：April 2016
• 年：2016
• 卷：100
• 期：7
• 页码：3023-3034
• 全文大小：4,853 KB
• 参考文献：Ansari N, Khodagholi F (2013) Molecular mechanism aspect of ER stress in Alzheimer’s disease: current approaches and future strategies. Curr Drug Targets 14(1):114–122CrossRef PubMed
  Baneyx F, Mujacic M (2004) Recombinant protein folding and misfolding in Escherichia coli. Nat Biotechnol 22(11):1399–1408. doi:10.​1038/​nbt1029 CrossRef PubMed
  Banhegyi G, Baumeister P, Benedetti A, Dong D, Fu Y, Lee AS, Li J, Mao C, Margittai E, Ni M, Paschen W, Piccirella S, Senesi S, Sitia R, Wang M, Yang W (2007) Endoplasmic reticulum stress. Ann N Y Acad Sci 1113:58–71. doi:10.​1196/​annals.​1391.​007 CrossRef PubMed
  Boilly B, Vercoutter-Edouart AS, Hondermarck H, Nurcombe V, Le Bourhis X (2000) FGF signals for cell proliferation and migration through different pathways. Cytokine Growth Factor Rev 11(4):295–302CrossRef PubMed
  Chen H, Tung YC, Li B, Iqbal K, Grundke-Iqbal I (2007) Trophic factors counteract elevated FGF-2-induced inhibition of adult neurogenesis. Neurobiol Aging 28(8):1148–1162. doi:10.​1016/​j.​neurobiolaging.​2006.​05.​036 CrossRef PubMed
  de Mena L, Cardo LF, Coto E, Miar A, Diaz M, Corao AI, Alonso B, Ribacoba R, Salvador C, Menendez M, Moris G, Alvarez V (2010) FGF20 rs12720208 SNP and microRNA-433 variation: no association with Parkinson’s disease in Spanish patients. Neurosci Lett 479(1):22–25. doi:10.​1016/​j.​neulet.​2010.​05.​019 CrossRef PubMed
  Fan H, Vitharana SN, Chen T, O’Keefe D, Middaugh CR (2007) Effects of pH and polyanions on the thermal stability of fibroblast growth factor 20. Mol Pharm 4(2):232–240. doi:10.​1021/​mp060097h CrossRef PubMed
  Foo JN, Tan LC, Liany H, Koh TH, Irwan ID, Ng YY, Ahmad-Annuar A, Au WL, Aung T, Chan AY, Chong SA, Chung SJ, Jung Y, Khor CC, Kim J, Lee J, Lim SY, Mok V, Prakash KM, Song K, Tai ES, Vithana EN, Wong TY, Tan EK, Liu J (2014) Analysis of non-synonymous-coding variants of Parkinson’s disease-related pathogenic and susceptibility genes in east Asian populations. Hum Mol Genet 23(14):3891–3897. doi:10.​1093/​hmg/​ddu086 CrossRef PubMed
  Garcia-Fruitos E, Vazquez E, Diez-Gil C, Corchero JL, Seras-Franzoso J, Ratera I, Veciana J, Villaverde A (2012) Bacterial inclusion bodies: making gold from waste. Trends Biotechnol 30(2):65–70. doi:10.​1016/​j.​tibtech.​2011.​09.​003 CrossRef PubMed
  Grothe C, Timmer M (2007) The physiological and pharmacological role of basic fibroblast growth factor in the dopaminergic nigrostriatal system. Brain Res Rev 54(1):80–91. doi:10.​1016/​j.​brainresrev.​2006.​12.​001 CrossRef PubMed
  Hotamisligil GS (2010) Endoplasmic reticulum stress and the inflammatory basis of metabolic disease. Cell 140(6):900–917. doi:10.​1016/​j.​cell.​2010.​02.​034 CrossRef PubMed PubMedCentral
  Jeffers M, McDonald WF, Chillakuru RA, Yang M, Nakase H, Deegler LL, Sylander ED, Rittman B, Bendele A, Sartor RB, Lichenstein HS (2002) A novel human fibroblast growth factor treats experimental intestinal inflammation. Gastroenterology 123(4):1151–1162CrossRef PubMed
  Jeffers M, Shimkets R, Prayaga S, Boldog F, Yang M, Burgess C, Fernandes E, Rittman B, Shimkets J, LaRochelle WJ, Lichenstein HS (2001) Identification of a novel human fibroblast growth factor and characterization of its role in oncogenesis. Cancer Res 61(7):3131–3138PubMed
  Kinsella TJ, Kunugi KA, Vielhuber KA, Potter DM, Fitzsimmons ME, Collins JM (1998) Preclinical evaluation of 5-iodo-2-pyrimidinone-2′-deoxyribose as a prodrug for 5-iodo-2′-deoxyuridine-mediated radiosensitization in mouse and human tissues. Clin Cancer Res : Off J Am Assoc Cancer Res 4(1):99–109
  Kirikoshi H, Sagara N, Saitoh T, Tanaka K, Sekihara H, Shiokawa K, Katoh M (2000) Molecular cloning and characterization of human FGF-20 on chromosome 8p21.3-p22. Biochem Biophys Res Commun 274(2):337–343. doi:10.​1006/​bbrc.​2000.​3142 CrossRef PubMed
  Kiyota T, Ingraham KL, Jacobsen MT, Xiong H, Ikezu T (2011) FGF2 gene transfer restores hippocampal functions in mouse models of Alzheimer’s disease and has therapeutic implications for neurocognitive disorders. Proc Natl Acad Sci U S A 108(49):E1339–E1348. doi:10.​1073/​pnas.​1102349108 CrossRef PubMed PubMedCentral
  Li Y, Basilico C, Mansukhani A (1994) Cell transformation by fibroblast growth factors can be suppressed by truncated fibroblast growth factor receptors. Mol Cell Biol 14(11):7660–7669CrossRef PubMed PubMedCentral
  Liu R, Gao M, Qiang GF, Zhang TT, Lan X, Ying J, Du GH (2009) The anti-amnesic effects of luteolin against amyloid beta(25–35) peptide-induced toxicity in mice involve the protection of neurovascular unit. Neuroscience 162(4):1232–1243. doi:10.​1016/​j.​neuroscience.​2009.​05.​009 CrossRef PubMed
  Ma ZG, Xu J, Liu TW (2015) Quantitative assessment of the association between fibroblast growth factor 20 rs1721100 C/G polymorphism and the risk of sporadic Parkinson’s diseases: a meta-analysis. Neurol Sci : Off J Ital Neurol Soc Ital Soc Clin Neurophysiol 36(1):47–51. doi:10.​1007/​s10072-014-1884-4 CrossRef
  Maclachlan T, Narayanan B, Gerlach VL, Smithson G, Gerwien RW, Folkerts O, Fey EG, Watkins B, Seed T, Alvarez E (2005) Human fibroblast growth factor 20 (FGF-20; CG53135-05): a novel cytoprotectant with radioprotective potential. Int J Radiat Biol 81(8):567–579. doi:10.​1080/​0955300050021109​1 CrossRef PubMed
  Ohmachi S, Mikami T, Konishi M (2003) Preferential neurotrophic activity of fibroblast growth factor-20 for dopaminergic neurons through fibroblast growth factor receptor-1c. Journal of Neuroscience Research
  Ohmachi S, Watanabe Y, Mikami T, Kusu N, Ibi T, Akaike A, Itoh N (2000) FGF-20, a novel neurotrophic factor, preferentially expressed in the substantia nigra pars compacta of rat brain. Biochem Biophys Res Commun 277(2):355–360. doi:10.​1006/​bbrc.​2000.​3675 CrossRef PubMed
  Ori A, Wilkinson MC, Fernig DG (2011) A systems biology approach for the investigation of the heparin/heparan sulfate interactome. J Biol Chem 286(22):19892–19904. doi:10.​1074/​jbc.​M111.​228114 CrossRef PubMed PubMedCentral
  Schlessinger J, Plotnikov AN, Ibrahimi OA, Eliseenkova AV, Yeh BK, Yayon A, Linhardt RJ, Mohammadi M (2000) Crystal structure of a ternary FGF-FGFR-heparin complex reveals a dual role for heparin in FGFR binding and dimerization. Mol Cell 6(3):743–750CrossRef PubMed
  Shimada H, Yoshimura N, Tsuji A, Kunisada T (2009) Differentiation of dopaminergic neurons from human embryonic stem cells: modulation of differentiation by FGF-20. J Biosci Bioeng 107(4):447–454. doi:10.​1016/​j.​jbiosc.​2008.​12.​013 CrossRef PubMed
  Sleeman IJ, Boshoff EL, Duty S (2012) Fibroblast growth factor-20 protects against dopamine neuron loss in vitro and provides functional protection in the 6-hydroxydopamine-lesioned rat model of Parkinson’s disease. Neuropharmacology 63(7):1268–1277. doi:10.​1016/​j.​neuropharm.​2012.​07.​029 CrossRef PubMed
  Song L, Huang Z, Chen Y, Li H, Jiang C, Li X (2014) High-efficiency production of bioactive recombinant human fibroblast growth factor 18 in Escherichia coli and its effects on hair follicle growth. Appl Microbiol Biotechnol 98(2):695–704. doi:10.​1007/​s00253-013-4929-3 CrossRef PubMed
  Sun C, Li Y, Taylor SE, Mao X, Wilkinson MC, Fernig DG (2015) HaloTag is an effective expression and solubilisation fusion partner for a range of fibroblast growth factors. PeerJ 3:e1060. doi:10.​7717/​peerj.​1060 CrossRef PubMed PubMedCentral
  Swietnicki W (2006) Folding aggregated proteins into functionally active forms. Curr Opin Biotechnol 17(4):367–372. doi:10.​1016/​j.​copbio.​2006.​05.​011 CrossRef PubMed
  Takagi Y, Takahashi J, Saiki H, Morizane A, Hayashi T, Kishi Y, Fukuda H, Okamoto Y, Koyanagi M, Ideguchi M, Hayashi H, Imazato T, Kawasaki H, Suemori H, Omachi S, Iida H, Itoh N, Nakatsuji N, Sasai Y, Hashimoto N (2005) Dopaminergic neurons generated from monkey embryonic stem cells function in a Parkinson primate model. J Clin Invest 115(1):102–109. doi:10.​1172/​JCI21137 CrossRef PubMed PubMedCentral
  van der Walt JM, Noureddine MA, Kittappa R, Hauser MA, Scott WK, McKay R, Zhang F, Stajich JM, Fujiwara K, Scott BL, Pericak-Vance MA, Vance JM, Martin ER (2004) Fibroblast growth factor 20 polymorphisms and haplotypes strongly influence risk of Parkinson disease. Am J Hum Genet 74(6):1121–1127. doi:10.​1086/​421052 CrossRef PubMed PubMedCentral
  Wang G, van der Walt JM, Mayhew G, Li YJ, Zuchner S, Scott WK, Martin ER, Vance JM (2008) Variation in the miRNA-433 binding site of FGF20 confers risk for Parkinson disease by overexpression of alpha-synuclein. Am J Hum Genet 82(2):283–289. doi:10.​1016/​j.​ajhg.​2007.​09.​021 CrossRef PubMed PubMedCentral
  Xian YF, Ip SP, Lin ZX, Mao QQ, Su ZR, Lai XP (2012) Protective effects of pinostrobin on beta-amyloid-induced neurotoxicity in PC12 cells. Cell Mol Neurobiol 32(8):1223–1230. doi:10.​1007/​s10571-012-9847-x CrossRef PubMed
  Xu X, Wang N, Xu H, Xie A, Jiang H, Xie J (2013) Fibroblast growth factor 20 polymorphism in sporadic Parkinson’s disease in northern Han Chinese. J Clin Neurosci : Off J Neurosurgical Soc Australasia 20(11):1588–1590. doi:10.​1016/​j.​jocn.​2013.​02.​023 CrossRef
  Zhang K, Kaufman RJ (2008) From endoplasmic-reticulum stress to the inflammatory response. Nature 454(7203):455–462. doi:10.​1038/​nature07203 CrossRef PubMed PubMedCentral
  Zhu R, Zhu Y, Liu X, He Z (2014) Fibroblast growth factor 20 (FGF20) gene polymorphism and risk of Parkinson’s disease: a meta-analysis. Neurol Sci : Off J Ital Neurol Soc Ital Soc Clin Neurophysiol 35(12):1889–1894. doi:10.​1007/​s10072-014-1853-y CrossRef
  
• 作者单位：Haishan Tian (1)
  Yang Zhao (1)
  Nazi Chen (1)
  Meiyu Wu (1)
  Weiyue Gong (1)
  Jie Zheng (1)
  David G. Fernig (1)
  Alois Jungbauer (1)
  Dezhong Wang (1)
  Xiaokun Li (1)
  Chao Jiang (1)
  
  1. School of Pharmaceutical Science, Wenzhou Medical University, Chashan College Park, Wenzhou, 325035, China
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Microbiology
  Microbial Genetics and Genomics
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0614

文摘

Fibroblast growth factor 20 (FGF20) has a wide range of biological activities; its expression is most pronounced in neural tissues where it has functions in development and neuroprotection. Given these activities, interest in the clinical applications of FGF20 is rising, which will lead to increasing demand for active recombinant human FGF20 (rhFGF20). To improve the production of rhFGF20, an artificial gene encoding fgf20 was cloned into pET3a and expressed in E. coli BL21(DE3)pLysS. By optimizing induction conditions, we successfully induced large amounts of insoluble rhFGF20. Following solubilization and refolding of the rhFGF20 from inclusion bodies, it was purified by HiTrap heparin affinity chromatography to a purity of over 96 % with a yield of 218 mg rhFGF20/100 g wet cells. The purified rhFGF20 could stimulate proliferation of both NIH 3T3 cells and PC-12 cells, measured by the MTT assay. In a model of Aβ25–35-induced apoptosis on PC-12 cells, rhFGF20 had a clear protective effect. RT-PCR and Western blot analysis of apoptosis-related genes and proteins revealed that the FGF20-derived protective mechanism was likely due to the relief of endoplasmic reticulum stress (ER stress). In conclusion, the approach described here may be a better means to produce active rhFGF20 in good quantity, thereby allowing for its future pharmacological and clinical use.