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Genomics & informatics v.4 no.3, 2006년, pp.133 - 136  

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  );
  • 초록

    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.


  • 주제어

    his-tagged protein .   nickel-coated glass slides .   protein chip .   unfolding.  

  • 참고문헌 (12)

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