2012 Fiscal Year Final Research Report
Obesity and oxidative stress: involvement of SOD in the physiology of metabolic syndrome and atherosclerosis
| Project/Area Number |
22590563
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Hygiene
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| Research Institution | Hyogo University of Health Sciences |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
KOREKANE Atsuko 兵庫医療大学, 薬学部, 助教 (80461169)
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| 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)
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| Project Period (FY) |
2010 – 2012
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| Keywords | 予防医学 |
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| 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.
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