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Expression and purification of two alternative peptides for mechano-growth factor in Escherichia coli

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Abstract

Two genes, MGFc 40 and MGFc 24, encoding different E peptides of mechano-growth factor (MGF), were obtained by a four-step PCR strategy and subcloned into pRSETC and pGEX-6p-1. Recombinant MGFc40 protein (4 mg l−1) was expressed and purified by affinity chromatography using His60 Ni Superflow. Recombinant MGFc24 protein was purified using a glutathione-Sepharose 4B column. After enzymatic cleavage of the GST-tail, 1 mg MGFc24 protein l−1 was obtained. MGFc40 and MGFc24, which are involved in proliferation and differentiation, stimulated cell proliferation and inhibited cell differentiation of C2C12 cells.

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References

  • Ates K, Yang SY, Orrell RW, Sinanan AC, Simons P, Solomon A, Beech S, Goldspink G, Lewis MP (2007) The IGF-I splice variant MGF increases progenitor cells in ALS, dystrophic, and normal muscle. FEBS Lett 581:2727–2732

    Article  PubMed  CAS  Google Scholar 

  • Carpenter V, Matthews K, Devlin G, Stuart S, Jensen J, Conaglen J, Jeanplong F, Goldspink P, Yang SY, Goldspink G, Bass J, McMahon C (2008) Mechano-growth factor reduces loss of cardiac function in acute myocardial infarction. Heart Lung Circ 17:33–39

    Article  PubMed  Google Scholar 

  • Dai Z, Wu F, Yeung EW, Li Y (2010) IGF-IEc expression, regulation and biological function in different tissues. Growth Horm IGF Res 20:275–281

    Article  PubMed  CAS  Google Scholar 

  • Dluzniewska J, Sarnowska A, Beresewicz M, Johnson I, Srai SK, Ramesh B, Goldspink G, Gorecki DC, Zabtocka B (2005) A strong neuroprotective effect of the autonomous C-terminal peptide of IGF-1 Ec (MGF) in brain ischemia. FASEB J 19:1896–1898

    PubMed  CAS  Google Scholar 

  • Goldspink G (2005) Mechanical signals, IGF-I gene splicing, and muscle adaptation. Physiology 20:232–238

    Article  PubMed  CAS  Google Scholar 

  • Goldspink G (2006) Impairment of IGF-I gene splicing and MGF expression associated with muscle wasting. Int J Biochem Cell Biol 38:481–489

    Article  PubMed  CAS  Google Scholar 

  • Hill M, Goldspink G (2003) Expression and splicing of the insulin-like growth factor gene in rodent muscle is associated with muscle satellite (stem) cell activation following local tissue damage. J Physiol 549:409–418

    Article  PubMed  CAS  Google Scholar 

  • Kuznetsova TV, Schulga AA, Wulfson AN, Keruchenko JS, Ermolyuk YS, Keruchenko ID, Tikhonov RV, Lisitskaya KV, Makarov AA, Chobotova K, Khomenkov VG, Khotchenkov VP, Popov VO, Kirpichnikov MP, Shevelev AB (2008) Producing human mechano growth factor(MGF) in Escherichia coli. Protein Expr Purif 58:70–77

    Article  PubMed  CAS  Google Scholar 

  • Mills P, Dominique JC, Lafreniere JF, Bouchentouf M, Tremblay JP (2007) A synthetic mechano growth factor E Peptide enhances myogenic precursor cell transplantation success. Am J Transpl 7:2247–2259

    Article  CAS  Google Scholar 

  • Osses N, Brandan E (2002) ECM is required for skeletal muscle differentiation independently of muscle regulatory factor expression. Am J Physiol Cell Physiol 282:83–94

    Google Scholar 

  • Owino V, Yang SY, Goldspink G (2001) Age-related loss of skeletal muscle function and the inability to express the autocrine form of insulin-like growth factor-1 (MGF) in response to mechanical overload. FEBS Lett 505:259–263

    Article  PubMed  CAS  Google Scholar 

  • Riddoch-Contreras J, Yang SY, Dick JR, Goldspink G, Orrell RW, Greensmith L (2009) Mechano-growth factor, an IGF-I splice variant, rescues motoneurons and improves muscle function in SOD1(G93A) mice. Exp Neurol 215:281–289

    Article  PubMed  CAS  Google Scholar 

  • Yang SY, Goldspink G (2002) Different roles of the IGF-I Ec peptide (MGF) and mature IGF-I in myoblast proliferation and differentiation. FEBS Lett 522:156–160

    Article  PubMed  CAS  Google Scholar 

  • Yang S, Alnaqeeb M, Simpson H (1996) Cloning and characterization of an IGF-I isoform expressed in skeletal muscle subjected to stretch. J Muscle Res Cell Motil 17:487–495

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (No. 31040018). This research project was supported by the Shanxi Scholarship Council of China (2010-2012), Natural Sciences Foundation of Shanxi (2009021035-2) and International technical cooperation projects of Shanxi (2011081058).

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Correspondence to Li Song.

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Song, L., Zhu, WS., Dong, JL. et al. Expression and purification of two alternative peptides for mechano-growth factor in Escherichia coli . Biotechnol Lett 34, 231–237 (2012). https://doi.org/10.1007/s10529-011-0762-x

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  • DOI: https://doi.org/10.1007/s10529-011-0762-x

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