Abstract
Fibroblast growth factor (FGF) 13, a member of the FGF11 subfamily, is a kind of intracrine protein similar to other family members including FGF11, FGF12, and FGF14. Unlike classical FGF, FGF13 exerts its bioactivities independent of fibroblast growth factor receptors (FGFRs). However, the effect of exogenous administration of FGF13 still remains further investigated. In the present study, we established an Escherichia coli expression system for the large-scale production of FGF13 and then obtained two isoform proteins including recombinant human FGF13A (rhFGF13A) and rhFGF13B with a purity greater than 90% by column chromatography, respectively. Otherwise, soluble analysis indicated that both rhFGF13A and rhFGF13B expressed in E. coli BL21 (DE3) pLysS were soluble. Furthermore, cellular-based experiments demonstrated that rhFGF13A, rather than rhFGF13B, could promote the proliferation of NIH3T3 cells in the presence of heparin. Mechanistically, the mitogenic effect of FGF13 was mediated by activation of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK), but not p38. Moreover, blockage of FGFRs also significantly attenuated the mitogenic effects of rhFGF13A, implying that FGFRs are still related to FGF13. Thus, our research shows that exogenous FGF13 can act as secreted FGF to participate in cell signal transmission and heparin is still required as an ancillary cofactor for the mitogenic effects of FGF13, which may help people to discover more potential functions of FGF13 in cell life activities.
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References
Abu-Amero KK, Kondkar AA, Alorainy IA, Khan AO, Al-Enazy LA, Oystreck DT, Bosley TM (2014) Xq26.3 microdeletion in a male with Wildervanck syndrome. Ophthalmic Genet 35(1):18–24. https://doi.org/10.3109/13816810.2013.766218
Beenken A, Mohammadi M (2009) The FGF family: biology, pathophysiology and therapy. Nat Rev Drug Discov 8(3):235–253. https://doi.org/10.1038/nrd2792
Berry D, Shriver Z, Venkataraman G, Sasisekharan R (2004) Quantitative assessment of FGF regulation by cell surface heparan sulfates. Biochem Biophys Res Commun 314(4):994–1000. https://doi.org/10.1016/j.bbrc.2003.12.188
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):295–302
Chen G, Liu Y, Goetz R, Fu L, Jayaraman S, Hu MC, Moe OW, Liang G, Li X, Mohammadi M (2018) Alpha-Klotho is a non-enzymatic molecular scaffold for FGF23 hormone signalling. Nature 553(7689):461–466. https://doi.org/10.1038/nature25451
Chen T, Gong W, Tian H, Wang H, Chu S, Ma J, Yang H, Cheng J, Liu M, Li X, Jiang C (2017) Fibroblast growth factor 18 promotes proliferation and migration of H460 cells via the ERK and p38 signaling pathways. Oncol Rep 37(2):1235–1242. https://doi.org/10.3892/or.2016.5301
Cheng J, Fang Z, Yang H, Li Y, Tian H, Gong W, Chen T, Liu M, Li X, Jiang C (2017) High-yield of biologically active recombinant human fibroblast growth factor-16 in E. coli and its mechanism of proliferation in NCL-H460 cells. Prep Biochem Biotechnol 47(7):720–729. https://doi.org/10.1080/10826068.2017.1315599
DeStefano GM, Fantauzzo KA, Petukhova L, Kurban M, Tadin-Strapps M, Levy B, Warburton D, Cirulli ET, Han Y, Sun X, Shen Y, Shirazi M, Jobanputra V, Cepeda-Valdes R, Cesar Salas-Alanis J, Christiano AM (2013) Position effect on FGF13 associated with X-linked congenital generalized hypertrichosis. Proc Natl Acad Sci U S A 110(19):7790–7795. https://doi.org/10.1073/pnas.1216412110
Gecz J, Baker E, Donnelly A, Ming JE, McDonald-McGinn DM, Spinner NB, Zackai EH, Sutherland GR, Mulley JC (1999) Fibroblast growth factor homologous factor 2 (FHF2): gene structure, expression and mapping to the Borjeson-Forssman-Lehmann syndrome region in Xq26 delineated by a duplication breakpoint in a BFLS-like patient. Hum Genet 104(1):56–63
Goetz R, Dover K, Laezza F, Shtraizent N, Huang X, Tchetchik D, Eliseenkova AV, Xu CF, Neubert TA, Ornitz DM, Goldfarb M, Mohammadi M (2009) Crystal structure of a fibroblast growth factor homologous factor (FHF) defines a conserved surface on FHFs for binding and modulation of voltage-gated sodium channels. J Biol Chem 284(26):17883–17896. https://doi.org/10.1074/jbc.M109.001842
Goldfarb M (2005) Fibroblast growth factor homologous factors: evolution, structure, and function. Cytokine Growth Factor Rev 16(2):215–220. https://doi.org/10.1016/j.cytogfr.2005.02.002
Goldfarb M, Schoorlemmer J, Williams A, Diwakar S, Wang Q, Huang X, Giza J, Tchetchik D, Kelley K, Vega A, Matthews G, Rossi P, Ornitz DM, D'Angelo E (2007) Fibroblast growth factor homologous factors control neuronal excitability through modulation of voltage-gated sodium channels. Neuron 55(3):449–463. https://doi.org/10.1016/j.neuron.2007.07.006
Hennessey JA, Wei EQ, Pitt GS (2013) Fibroblast growth factor homologous factors modulate cardiac calcium channels. Circ Res 113(4):381–388. https://doi.org/10.1161/CIRCRESAHA.113.301215
Johnson GL, Lapadat R (2002) Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science 298(5600):1911–1912. https://doi.org/10.1126/science.1072682
Kato T, Yamada A, Ikehata M, Yoshida Y, Sasa K, Morimura N, Sakashita A, Iijima T, Chikazu D, Ogata H, Kamijo R (2018) FGF-2 suppresses expression of nephronectin via JNK and PI3K pathways. FEBS open bio 8(5):836–842. https://doi.org/10.1002/2211-5463.12421
Kettunen P, Furmanek T, Chaulagain R, Kvinnsland IH, Luukko K (2011) Developmentally regulated expression of intracellular Fgf11-13, hormone-like Fgf15 and canonical Fgf16, -17 and -20 mRNAs in the developing mouse molar tooth. Acta Odontol Scand 69(6):360–366. https://doi.org/10.3109/00016357.2011.568968
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):7660–7669
Lu H, Shi X, Wu G, Zhu J, Song C, Zhang Q, Yang G (2015) FGF13 regulates proliferation and differentiation of skeletal muscle by down-regulating Spry1. Cell Prolif 48(5):550–560. https://doi.org/10.1111/cpr.12200
Meloche S, Pouyssegur J (2007) The ERK1/2 mitogen-activated protein kinase pathway as a master regulator of the G1- to S-phase transition. Oncogene 26(22):3227–3239. https://doi.org/10.1038/sj.onc.1210414
Mohammadi M, Olsen SK, Ibrahimi OA (2005) Structural basis for fibroblast growth factor receptor activation. Cytokine Growth Factor Rev 16(2):107–137. https://doi.org/10.1016/j.cytogfr.2005.01.008
Olsen SK, Garbi M, Zampieri N, Eliseenkova AV, Ornitz DM, Goldfarb M, Mohammadi M (2003) Fibroblast growth factor (FGF) homologous factors share structural but not functional homology with FGFs. J Biol Chem 278(36):34226–34236. https://doi.org/10.1074/jbc.M303183200
Ornitz DM (2000) FGFs, heparan sulfate and FGFRs: complex interactions essential for development. Bioessays 22(2):108–112. https://doi.org/10.1002/(sici)1521-1878(200002)22:2<108::aid-bies2>3.0.co;2-m
Rubinfeld H, Seger R (2005) The ERK cascade: a prototype of MAPK signaling. Mol Biotechnol 31(2):151–174. https://doi.org/10.1385/mb:31:2:151
Schoorlemmer J, Goldfarb M (2001) Fibroblast growth factor homologous factors are intracellular signaling proteins. Curr Biol : CB 11(10):793–797
Smallwood PM, Munoz-Sanjuan I, Tong P, Macke JP, Hendry SH, Gilbert DJ, Copeland NG, Jenkins NA, Nathans J (1996) Fibroblast growth factor (FGF) homologous factors: new members of the FGF family implicated in nervous system development. Proc Natl Acad Sci U S A 93(18):9850–9857
Wang C, Hennessey JA, Kirkton RD, Wang C, Graham V, Puranam RS, Rosenberg PB, Bursac N, Pitt GS (2011) Fibroblast growth factor homologous factor 13 regulates Na+ channels and conduction velocity in murine hearts. Circ Res 109(7):775–782. https://doi.org/10.1161/CIRCRESAHA.111.247957
Weston CR, Davis RJ (2007) The JNK signal transduction pathway. Curr Opin Cell Biol 19(2):142–149. https://doi.org/10.1016/j.ceb.2007.02.001
Wozniak DF, Xiao M, Xu L, Yamada KA, Ornitz DM (2007) Impaired spatial learning and defective theta burst induced LTP in mice lacking fibroblast growth factor 14. Neurobiol Dis 26(1):14–26. https://doi.org/10.1016/j.nbd.2006.11.014
Wu Q-F, Yang L, Li S, Wang Q, Yuan X-B, Gao X, Bao L, Zhang X (2012) Fibroblast growth factor 13 is a microtubule-stabilizing protein regulating neuronal polarization and migration. Cell 149(7):1549–1564. https://doi.org/10.1016/j.cell.2012.04.046
Zhang X, Bao L, Yang L, Wu Q, Li S (2012) Roles of intracellular fibroblast growth factors in neural development and functions. Sci China Life Sci 55(12):1038–1044. https://doi.org/10.1007/s11427-012-4412-x
Zhang X, Ibrahimi OA, Olsen SK, Umemori H, Mohammadi M, Ornitz DM (2006) Receptor specificity of the fibroblast growth factor family. The complete mammalian FGF family. J Biol Chem 281(23):15694–15700. https://doi.org/10.1074/jbc.M601252200
Zheng J, Wang S, Yang H, Chen Z, Huang S, Zhao T, Pan X, Fernig DG, Jiang C, Li X, Tian H (2017) Large-scale expression, purification of bioactive recombinant human FGF6 in E. coli and the mechanisms of its myocardial protection. Int J Pept Res Ther 24(1):105–115. https://doi.org/10.1007/s10989-017-9592-6
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This work was supported by the Opening Project of Zhejiang Provincial TOP Key Discipline of Pharmaceutical Sciences (No.201720).
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Lin, H., Lu, P., Zhou, M. et al. Purification of recombinant human fibroblast growth factor 13 in E. coli and its molecular mechanism of mitogenesis. Appl Microbiol Biotechnol 103, 7017–7027 (2019). https://doi.org/10.1007/s00253-019-09973-y
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DOI: https://doi.org/10.1007/s00253-019-09973-y