Journal of the Korean Magnetic Resonance Society (한국자기공명학회논문지)

Korean Magnetic Resonance Society (한국자기공명학회)
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Backbone NMR assignments of recombinant glucagon at pH 6.0

  • Sung-Hee Lee (Department of Biotechnology and Research Institute for Biomedical & Health Science, College of Biomedical & Health Science, Konkuk University) ;
  • Seong-Eun Park (Department of Biotechnology and Research Institute for Biomedical & Health Science, College of Biomedical & Health Science, Konkuk University) ;
  • Kyeong-Hyeon Yoon (Department of Biotechnology and Research Institute for Biomedical & Health Science, College of Biomedical & Health Science, Konkuk University) ;
  • Young-Ho Lee (Biopharmaceutical Research Center, Korea Basic Science Institute) ;
  • Eun-Hee Kim (Biopharmaceutical Research Center, Korea Basic Science Institute) ;
  • Hae-Kap Cheong (Biopharmaceutical Research Center, Korea Basic Science Institute) ;
  • Ji-Hun Kim (College of Pharmacy, Chungbuk National University) ;
  • Hyung-Sik Won (Department of Biotechnology and Research Institute for Biomedical & Health Science, College of Biomedical & Health Science, Konkuk University)
  • Received : 2024.11.25
  • Accepted : 2024.12.03
  • Published : 2024.12.20
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Abstract

The peptide hormone glucagon has served as both a biopharmaceutical agent for clinical use and a model peptide forming amyloid fibrils. As we recently established a recombinant production of glucagon, its backbone NMR assignments at neutral pHs were conducted in the present study. However, as the NMR spectra showed severe line broadening and poor dispersion at pHs 7.5 and 7.0, complete backbone NMR assignments could be obtained at pH 6.0. Interestingly, CSI and TALOS predictions using the assigned chemical shift values indicated different probabilities of secondary structure. These results suggest that glucagon would adopt an unusual or dynamically fluctuating conformation in solution. Therefore, detailed conformation and molecular dynamics of glucagon would be worthy of investigation, for which the present results provide an experimental basis.

Keywords

Acknowledgement

This paper was written as part of Konkuk University's research support program for its faculty on sabbatical leave in 2023 (H.-S.W.). This research was supported by the National Research Foundation of Korea (NRF) grants funded by the Korea governments, Ministry of Science and ICT (MSIT) (No. RS-2023-00221332, H.-S.W.; NRF2022R1A2C1011793, Y.-H.L.) and Ministry of Education (MOE) ("Regional Innovation Strategy (RIS)" No. 2021RIS-001, H.-S.W.). The use of NMR equipment was supported by the Korea Basic Science Institute (KBSI; Ochang, Korea) under the R&D program (Project No. A412550) supervised by the MSIT, Korea, and also by KBSI funds (Y.-H.L.; No. A439200, A423100, A412580, A423310, A412550, and A412120).

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