Journal of the Korean Magnetic Resonance Society (한국자기공명학회논문지)

Korean Magnetic Resonance Society (한국자기공명학회)
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Expression and Purification of the Helicase-like Subdomains, H1 and H23, of Reverse Gyrase from A. fulgidus for Heteronuclear NMR study

  • Kwon, Mun-Young (Department of Chemistry and RINS, Gyeongsang National University) ;
  • Seo, Yeo-Jin (Department of Chemistry and RINS, Gyeongsang National University) ;
  • Lee, Yeon-Mi (Department of Chemistry and RINS, Gyeongsang National University) ;
  • Lee, Ae-Ree (Department of Chemistry and RINS, Gyeongsang National University) ;
  • Lee, Joon-Hwa (Department of Chemistry and RINS, Gyeongsang National University)
  • Received : 2015.08.08
  • Accepted : 2015.08.25
  • Published : 2015.10.10
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Abstract

Reverse gyrase is a hyperthermophile specific protein which introduces positive supercoils into DNA molecules. Reverse gyrase consists of an N-terminal helicase-like domain and a C-terminal topoisomerase domain. The helicase-like domain shares the three-dimensional structure with two tandem RecA-folds (H1 and H2), in which the subdomain H2 is interrupted by the latch domain (H3). To understand the physical property of the hyperthermophile-specific protein, two subdomains af_H1 and af_H23 have been cloned into E. coli expression vector, pET28a. The $^{15}N$-labeled af_H1 and af_H23 proteins were expressed and purified for heteronuclear NMR study. The af_H1 protein exhibits the well-dispersion of amide signals in its $^1H/^{15}N$-HSQC spectra and thus further NMR study continues to be progressed.

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