Regulation of heme oxygenase-2 activity elucidated by the analysis of structural dynamics

• Project/Area Number: 26840040
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Functional biochemistry
• Research Institution: Yokohama City University (2015-2016); Suntory Foundation for Life Sciences (2014)
• Principal Investigator: Furukawa Ayako 横浜市立大学, 生命医科学研究科, 特任助教 (90453050)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2014: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 動的構造解析 / 酵素 / タンパク質の動的構造解析 / 酵素の調節機構 / 蛋白質の動的構造解析

Outline of Final Research Achievements

Heme oxygenase-2 (HO-2) is composed of a structured region including the heme-binding site and a C-terminal disordered region.However, the C-terminal region's role in the activity is elusive and is difficult to explain by the crystal structure alone because it is disordered. We hypothesized that the C-terminal region transiently interacts with the structured region to regulate the structural dynamics responsible for the activity. To test this hypothesis, we have examined the enzyme activity and structural dynamics of two HO-2 constructs with (long HO-2) and without (short HO-2) the C-terminal region. Enzyme activity assay showed that long HO-2 had 1.5 times higher activity than short HO-2. To explain the difference in the enzyme activity, we performed PRE, R2 dispersion, and CLEANEX-PM experiments. The results of these experiments suggest that the change of the fluctuating mode by the interaction with the C-terminal disordered region regulates the HO-2 activity.

Research Products

• [Journal Article] Quantitative analysis of protein–ligand interactions by NMR (2016)
  • Author(s): Furukawa A., Konuma T., Yanaka S., Sugase K.,
  • Journal Title: Progress in Nuclear Magnetic Resonance Spectroscopy Volume: 96 Pages: 47-57
  • Peer Reviewed
• [Journal Article] Quantitative analysis of location- and sequence-dependent deamination by APOBEC3G using real-time NMR spectroscopy (2014)
  • Author(s): Furukawa, A., Sugase, K., Morishita, R., Nagata, T., Kodaki, T., Takaori-Kondo, A., Ryo, A., Katahira, M.
  • Journal Title: Angew. Chem. Int. Ed. Engl. Volume: 53 Issue: 9 Pages: 2349-2352
  • DOI: 10.1002/anie.201309940
  • NAID: 120005647511
  • Peer Reviewed / Open Access
• [Presentation] Heme oxygenase-2 activity regulated by the C-terminal intrinsically disordered (2014)
  • Author(s): Sugase K.
  • Organizer: The 11th Japan-Korea Bilateral Symposium on Biological NMR
  • Place of Presentation: 大阪大学 蛋白研
  • Year and Date: 2014-12-19
  • Invited
• [Presentation] Regulation of heme oxygenase-2 activity elucidated by the analysis of structural dynamics (2014)
  • Author(s): Furukawa A.,, Yamamoto T., Matsuki Y., Suematsu M., Sugase K.
  • Organizer: International Conference on Magnetic Resonance in Biological Systems
  • Place of Presentation: Dallas, USA
  • Year and Date: 2014-08-24 – 2014-08-29