한국자기공명학회논문지 (Journal of the Korean Magnetic Resonance Society)

한국자기공명학회 (Korean Magnetic Resonance Society)
권호 표지
DOI QR코드

DOI QR Code

Backbone NMR chemical shift assignment for the substrate binding domain of Escherichia coli HscA

  • Jin Hae Kim (Department of New Biology, Daegu Gyeongbuk Institute of Science and Technology (DGIST))
  • 투고 : 2024.06.19
  • 심사 : 2024.06.24
  • 발행 : 2024.06.20
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

HscA is a Hsp70-type chaperone protein that plays an essential role to mediate the iron-sulfur (Fe-S) cluster biogenesis mechanism in Escherichia coli. Like other Hsp70 chaperones, HscA is composed of two domains: the nucleotide binding domain (NBD), which can hydrolyze ATP and use its chemical energy to facilitate the Fe-S cluster transfer process, and the substrate binding domain (SBD), which directly interacts with the substrate, IscU, the scaffold protein of an Fe-S cluster. In the present work, we prepared the isolated SBD construct of HscA (HscA(SBD)) and conducted the solution-state nuclear magnetic resonance (NMR) experiments to have its backbone chemical shift assignment information. Due to low spectral quality of HscA(SBD), we obtained all the NMR data from the sample containing the peptide LPPVKIHC, the HscA-interaction motif of IscU, from which the chemical shift assignment could be done successfully. We expect that this information provides an important basis to execute detailed structural characterization of HscA and appreciate its interaction with IscU.

키워드

과제정보

We are grateful to Dr. Marco Tonelli and Dr. John Markley (Nuclear Magnetic Resonance Facility at Madison, University of Wisconsin-Madison, USA) for their generous help to conduct the NMR experiments. This research was supported by the National Research Foundation of Korea (NRF) funded by the Ministry of Science & ICT (NRF-2020R1I1A2074335).

참고문헌

  1. H. Beinert, R. H. Holm, and E. Munck, Science 277, 653(1997)
  2. S. Bandyopadhyay, K. Chandramouli, and M. K. Johnson, Biochem. Soc. Trans. 36, 1112 (2008)
  3. D. C. Johnson, D. R. Dean, A. D. Smith, and M. K. Johnson, Annu Rev Biochem. 74, 247 (2005)
  4. K. Chandramouli and M. K. Johnson, Biochemistry 45, 11087 (2006)
  5. F. Bonomi, S. Iametti, A. Morleo, D. Ta, and L. E. Vickery, Biochemistry 47, 12795 (2008)
  6. J. R. Cupp-Vickery, J. C. Peterson, D. T. Ta, and L. E. Vickery, J. Mol. Biol. 342, 1265 (2004)
  7. J. L. Markley, J. Kim, Z. Dai, et al. FEBS Lett. 587, 1172 (2013)
  8. T. L. Tapley, J. R. Cupp-Vickery, and L. E. Vickery Biochemistry 45, 8058 (2006)
  9. J. Kim, M. Tonelli, R. O. Frederick, D. C. F. Chow, and J. L. Markley, J. Biol. Chem. 287, 31406 (2012)
  10. W. Lee, M. Tonelli, and J. L. Markley, Bioinformatics 31, 1325 (2015)
  11. W. Lee, M. Rahimi, Y. Lee, and A. Chiu. Bioinformatics 37, 3041 (2021)