Journal of the Korean Magnetic Resonance Society (한국자기공명학회논문지)

Korean Magnetic Resonance Society (한국자기공명학회)
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Production and Amyloid fibril formation of tandem repeats of recombinant Yeast Prion like protein fragment

  • Kim, Yong-Ae (Department of Chemistry, Hankuk University of Foreign Studies) ;
  • Park, Jae-Joon (Department of Chemistry, Hankuk University of Foreign Studies) ;
  • Hwang, Jung-Hyun (Department of Chemistry, Hankuk University of Foreign Studies) ;
  • Park, Tae-Joon (Department of Chemistry, Hankuk University of Foreign Studies)
  • Received : 2011.11.30
  • Accepted : 2011.12.13
  • Published : 2011.12.20
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Abstract

Amyloid fibrils have long been known to be the well known ${\alpha}$-helix to ${\beta}$-sheet transition characterizing the conversion of cellular to scrapie forms of the prion protein. A very short sequence of Yeast prion-like protein, GNNQQNY (SupN), is responsible for aggregation that induces diseases. KSI-fused tandem repeats of SupN vector are constructed and used to express SupN peptide in Escherichia coli (E.Coli). A method for a production, purification, and cleavage of tandem repeats of recombinant isotopically enriched SupN in E. coli is described. This method yields as much as 20 mg/L of isotope-enriched fusion proteins in minimal media. Synthetic SupN peptides and $^{13}C$ Gly labeled SupN peptides are studied by Congo Red staining, Birefringence and transmission electron microscopy to characterize amyloid fibril formation. To get a better understanding of aggregation-structure relationship of 7 residues of Yeast prion-like protein, the change of a conformational structure will be studied by $^{13}C$ solid-state nmr spectroscopy as powder of both amorphous and fibrillar forms.

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References

  1. J.W. Kelly, Curr. Opi. Struct. Biol. 8, 101, (1998). https://doi.org/10.1016/S0959-440X(98)80016-X
  2. C.M. Dobson, Trends Biochem. Sci. 24, 329, (1999). https://doi.org/10.1016/S0968-0004(99)01445-0
  3. R.B. Wickner, Science 264, 566, (1994). https://doi.org/10.1126/science.7909170
  4. H. E. Sparrer, A. Santoso, F.C. Szoka, J.S. Weissman Science 289, 595, (2000). https://doi.org/10.1126/science.289.5479.595
  5. T. J. Park, S. Y. Im, J. S. Kim, Y. Kim, Proc. Biochem. 45, 682, (2010). https://doi.org/10.1016/j.procbio.2010.01.002
  6. J. S. Kim, T. J. Park, Y. Kim, J. Kor. Magn. Reson. 13, 96, (2009). https://doi.org/10.6564/JKMRS.2009.13.2.096
  7. M. Reaches, Y. Porant, E. Gazit, J. Biol. Chem. 277, 35475, (2002). https://doi.org/10.1074/jbc.M206039200
  8. W. Zou, D. Sheng, P. X. Fraser, N. R. Cashman, A. Chakrabartty, Eur. J. Biochem. 268, 4885, (2001). https://doi.org/10.1046/j.1432-1327.2001.02415.x
  9. R. Khurana, V. N. Uversky, L. Nielson, A. L. Fink, J. Biol. Chem. 276, 22715, (2001). https://doi.org/10.1074/jbc.M011499200
  10. X. Wu and K. W. Zilm, J. Mag. Res. A 111, 29, (1994).
  11. O.N. Antzutkin , R. Tycko, J. Chem. Phys. 110 , 2749, (1999). https://doi.org/10.1063/1.477876