Browse > Article
http://dx.doi.org/10.6564/JKMRS.2015.19.2.095

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)
Publication Information
Journal of the Korean Magnetic Resonance Society / v.19, no.2, 2015 , pp. 95-98 More about this Journal
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.
Keywords
NMR; reverse gyrase; subdomain; protein purification; helicase-like; hyperthermophile;
Citations & Related Records
Times Cited By KSCI : 3  (Citation Analysis)
연도 인용수 순위
1 C. De La Tour, C. Portemer, R. Huber, P. Forterre, and M. Duguet, J. Bacteriol. 173, 3921 (1991)   DOI
2 M. Heine and S. B. C. Chandra, J.Microbiol. 47, 229 (2009)   DOI
3 G. Perugino, A. Valenti, A. D'Amaro, M. Rossi, and M. Ciaramella, Biochem. Soc. Trans. 37, 69 (2009)   DOI
4 M. Nadal, Biochimie 89, 447 (2007)   DOI
5 Y. del Toro Duany, S. P. Jungblut, A. S. Schmidt, and D. Klostermeier, Nucleic Acids Res. 36, 5882 (2008)   DOI
6 A. C. Rodriguez and D. Stock, EMBO J. 21, 418 (2002)   DOI
7 B. Theissen, A. R. Karow, J. Kohler, A. Gubaev, and D. Klostermeier, Proc. Natl. Acad. Sci. USA 105, 548 (2008)   DOI
8 T. S. Hsieh and J. L. Plank J. Biol. Chem. 281, 5640 (2006)   DOI
9 F. Delaglio, S. Grzesiek, G. W. Vuister, G. Zhu, J. Pfeifer, and A. Bax, J. Biomol. NMR 6, 277 (1995)
10 T. D. Goddard and D. G. Kneller, SPARKY 3. University of California, San Francisco, CA. (2003)
11 J.-Y. Suh, T.- K. Yu, Y.- J. Yun, and K. O. Lee, J. Korean Magn. Reson. Soc. 18, 1 (2014)
12 S. J. Park, J. Korean Magn. Reson. Soc. 18, 47 (2014)   DOI