Obesity and oxidative stress: involvement of SOD in the physiology of metabolic syndrome and atherosclerosis

• Project/Area Number: 22590563
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Hygiene
• Research Institution: Hyogo University of Health Sciences
• Principal Investigator: OOKAWARA Tomomi 兵庫医療大学, 薬学部, 教授 (50330452)
• Co-Investigator(Kenkyū-buntansha): KOREKANE Atsuko 兵庫医療大学, 薬学部, 助教 (80461169)
• Co-Investigator(Renkei-kenkyūsha): FUJIWARA Noriko 兵庫医科大学, 医学部, 准教授 (10368532) ; SUZUKI Keiichiro 兵庫医科大学, 医学部, 教授 (70221322)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2012: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2011: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2010: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 予防医学 / 酸化ストレス / 細胞外型スーパーオキシドジスムターゼ / 動脈硬化 / N結合型糖鎖修飾 / 高血圧症 / 脂肪前駆細胞 / 肥満 / 活性酸素 / スーパーオキシドジスムターゼ / メタボリック症候群 / N-結合型糖鎖

Research Abstract

Recombinant extracellular superoxide dismutases were purified and the heparin affinity was investigated. 104 micrograms of the wild-type human enzyme was purified from 473mL of cultured medium of recombinant EC-SOD expressing cell line HECY6-7 through three steps of chromatography, heparin-affinity, hydroxyapatite, and gel-filtration. Removing the N-glycan reduced reactivity of ConA, and increased both of specific activity and heparin affinity of the enzyme. Two mutation studies at heparin binding domain (substitution of amino acid, Glu237Lys and doubling basic amino acid cluster, KERKKRRRESEC => KERKKRRRESECKEKKRRRESEC) gave rise to higher heparin binding EC-SOD. Another two mutation that moved the position of N-glycan (N121Q/Q207N and N121Q/S234N) affected the heparin affinity nature of the enzyme. These results suggest that the structure and position of N-glycan play an important role in the function and localization of EC-SOD.

Research Products

• [Journal Article] Structural switching of Cu,Zn-superoxide dismutases at loop VI: insights from the crystal structure of 2-mercaptoethanol-modified enzyme. (2012)
  • Author(s): Ihara K, Fujiwara N, Yamaguchi Y, Torigoe H, Wakatsuki S, Taniguchi N, Suzuki K.
  • Journal Title: Biosci Rep Volume: 32(6) Issue: 6 Pages: 539-48
  • DOI: 10.1042/bsr20120029
  • Peer Reviewed
• [Journal Article] Alterations in renal iron metabolism caused by a copper/zinc-superoxide dismutase deficiency. (2012)
  • Author(s): Yoshihara D, Fujiwara N, Kato S, Sakiyama H, Eguchi H, Suzuki K.
  • Journal Title: Free Radic Res Volume: 46(6) Issue: 6 Pages: 750-7
  • DOI: 10.3109/10715762.2012.673223
  • Peer Reviewed
• [Journal Article] N-Glycosylation profiling of recombinant mouse extracellular superoxide dismutase produced in Chinese hamster ovary cells (2011)
  • Author(s): Korekane H
  • Journal Title: Glycoconj J Volume: 28 Issue: 3-4 Pages: 183-196
  • DOI: 10.1007/s10719-011-9333-6
  • Peer Reviewed
• [Journal Article] Voluntary exercise attenuates obesity-associated inflammation through ghrelin expressed in macrophages (2011)
  • Author(s): Kizaki, T., Maegawa, T., Sakurai, T., Ogasawara, J., Ookawara, T., Oh-ishi, S., Izawa, T., Haga, S. & Ohno, H.
  • Journal Title: Biochem. Biophys. Res. Commun Volume: 413 Issue: 3 Pages: 454-459
  • DOI: 10.1016/j.bbrc.2011.08.117
  • Peer Reviewed
• [Journal Article] Hydrogen peroxide enhances LPS-induced nitric oxide production via the expression of interferon beta in BV-2 microglial cells. (2011)
  • Author(s): Eguchi H, Fujiwara N, Sakiyama H, Yoshihara D, Suzuki K.
  • Journal Title: Neurosci Lett Volume: 494(1) Issue: 1 Pages: 29-33
  • DOI: 10.1016/j.neulet.2011.02.047
  • Peer Reviewed
• [Journal Article] N-Glycosylation profiling of recombinant mouse extracellular superoxide dismutase produced in Chinese hamster ovary cells (2011)
  • Author(s): Hiroaki Korekane・Atsuko Korekane
  • Journal Title: Glycoconjugate Journal Volume: 28(In press)
  • Peer Reviewed