Journal of the Korean Magnetic Resonance Society (한국자기공명학회논문지)

Korean Magnetic Resonance Society (한국자기공명학회)
권호 표지
DOI QR코드

DOI QR Code

Characterization of pH-dependent structural properties of hydrolase PncA using NMR

  • Yi, Jong-Jae (College of pharmacy and Institute of Pharmaceutical Sciences, CHA University) ;
  • Kim, Won-Je (College of pharmacy, Seoul National University) ;
  • Rhee, Jin-Kyu (Department of Food Science and Engineering, Ewha Womans University) ;
  • Lim, Jongsoo (Discovery Technology Team, Dong-A ST Research Institute) ;
  • Lee, Bong-Jin (College of pharmacy, Seoul National University) ;
  • Son, Woo Sung (College of pharmacy and Institute of Pharmaceutical Sciences, CHA University)
  • Received : 2018.12.06
  • Accepted : 2018.12.17
  • Published : 2018.12.20
Download PDF
⟨ Previous Next ⟩

Abstract

Catalytic enzyme Pyrazinamidase (PncA) from Mycobacterium tuberculosis can hydrolyze substrate pyrazinamide (PZA) to pyrazoic acid (POA) as active form of compound. Using NMR spectroscopy, pH-dependent catalytic properties were monitored including metal binding mode during converting PZA to POA. There seems to be a conformational change through zinc binding in active site from the perturbation of peak intensities in series of 2D HSQC spectra the conformation changes through zinc binding.

Keywords

JGGMB2_2018_v22n4_144_f0001.png 이미지

Figure 1. 2D HSQC spectra for pH titration of PncA. 2D HSQC spectra were plotted in various pH (colored by black (pH 5.3), red (pH 5.6), green (pH 5.9), blue (pH 6.1) and magenta (pH 6.7).

JGGMB2_2018_v22n4_144_f0002.png 이미지

Figure 2. 2D HSQC spectra of zinc titration of PncA. One equivalent of zinc (red) was added to native PncA sample in pH 6.7 (black).

JGGMB2_2018_v22n4_144_f0003.png 이미지

Figure 3. Analysis of 1H-15N HSQC spectra of PncA with and without zinc ion at 313 K, pH 6.7. Chemical shift differences (top) for all clearly detectable peaks and relative peak intensity (bottom) of two HSQC spectra with and without zinc ion were shown.

JGGMB2_2018_v22n4_144_f0004.png 이미지

Figure 4. Active site comparison of PncA. Active site residues of P. horikoshii PncA was colored by orange and that of M. tuberculosis by yellow. Histidine 57 was colored by red, substrate PZA by cyan and zinc ion by blue.

References

  1. Y. Zhang and D. Mitchison, Int. J. Tuberc. Lung. Dis. 7, 6 (2003)
  2. W. Kim, J. Rhee, J. Yi, B. Lee, and W. S. Son, J. Kor. Magn. Reson. Soc. 18, 36 (2014) https://doi.org/10.6564/JKMRS.2014.18.1.036
  3. W. Kim, J. Rhee, J. Yi, B. Lee, and W. S. Son, J. Kor. Magn. Reson. Soc. 21, 85 (2017) https://doi.org/10.6564/JKMRS.2017.21.3.085
  4. N. Lemaitre, I. Callebaut, F. Frenois, V. Jarlier, and W. Sougakoff, Biochem. J. 353, 453 (2001)
  5. World Health Organization, WHO report 2006 (2006)
  6. X. Du, W. Wang, R. Kim, H. Yakota, H. Nguyen, and S. Kim, Biochemistry 40, 14166 (2001) https://doi.org/10.1021/bi0115479
  7. A. Somoskovi, L. M. Parsons, and M. Salfinger, Respir. Res. 2, 164 (2001) https://doi.org/10.1186/rr54
  8. J. Yi, J. K. Yoo, J. K. Kim, and W. S. Son, J. Kor. Magn. Reson. Soc. 17, 30 (2013) https://doi.org/10.6564/JKMRS.2013.17.1.030
  9. G. M. Morris, R. huey, W. Lindstrom, M. F.Sanner, R. K. Belew, D. S. Goodsell, and A. J. Olson, J. Comput. Chem. 30, 2785 (2009) https://doi.org/10.1002/jcc.21256
  10. R. Huey, G. M. Morris, A. J. Olson, and D. S. Goodsell, J. Comput. Chem. 28, 1145 (2007) https://doi.org/10.1002/jcc.20634
  11. F. A. Mulder, D. Schipper, R. Bott, and R. Boelens, J. Mol. Biol. 292, 111 (1999) https://doi.org/10.1006/jmbi.1999.3034
  12. T. R. Jahn, M. J. Parker, S. W. Homans, and S. E. Radford, Nat. Struct. Mol. Biol. 13, 195 (2006) https://doi.org/10.1038/nsmb1058