Genomics & Informatics

Korea Genome Organization (한국유전체학회)
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Unfolded Histidine-Tagged Protein is Immobilized to Nitrilotriacetic Acid-Nickel Beads, But Not the Nickel-Coated Glass Slide

  • Cho Min-Ho (Department of Molecular Biology & Institute of Nanosensor and Biotechnology, BK21 Graduate Program for RNA biology, Dankook University) ;
  • Ahn Sun-Young (Department of Molecular Biology & Institute of Nanosensor and Biotechnology, BK21 Graduate Program for RNA biology, Dankook University) ;
  • Park Heon-Yong (Department of Molecular Biology & Institute of Nanosensor and Biotechnology, BK21 Graduate Program for RNA biology, Dankook University)
  • Published : 2006.09.01
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Abstract

The adsorption of proteins on the surface of glass slides is essential for construction of protein chips. Previously, we prepared a nickel-coated plate by the spin-coating method for immobilization of His-tagged proteins. In order to know whether the structural factor is responsible for the immobilization of His-tagged proteins to the nickel-coated glass slide, we executed a series of experiments. First we purified a His-tagged protein after expressing the vector in E. coli BL21 (DE3). Then we obtained the unfolding curve for the His-tagged protein by using guanidine hydrochloride. Fractions unfolded were monitored by internal fluorescence spectroscopy. The ${\Delta}G_{H20}$ for unfolding was $2.27kcal/mol{/pm}0.52$. Then we tested if unfolded His-tagged proteins can be adsorbed to the nickel-coated plate, comparing with $Ni^{2+}-NTA$ (nitrilotriacetic acid) beads. Whereas unfolded His-tagged proteins were adsorbed to $Ni^{2+}-NTA$ beads, they did not bind to the nickel-coated plate. In conclusion, a structural factor is likely to be an important factor for constructing the protein chips, when His-tagged proteins will immobilize to the nickel-coated slides.

Keywords

References

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