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Conformational change in FALS-linked mutant Cu/Zn-SOD analyzed by monoclonal antibodies

Research Project

Project/Area Number 17500242
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Nerve anatomy/Neuropathology
Research InstitutionHyogo College of Medicine

Principal Investigator

FUJIWARA Noriko  Hyogo College of Medicine, Faculty of Medicine, Assistant Professor, 医学部, 講師 (10368532)

Co-Investigator(Kenkyū-buntansha) SUZUKI Keiichiro  Hyogo College of Medicine, Faculty of Medicine, Professor, 医学部, 教授 (70221322)
OOKAWARA Tomomi  Hyogo College of Medicine, Faculty of Medicine, Assistant Professor, 医学部, 講師 (50330452)
EGUCHI Hironobu  Hyogo College of Medicine, Faculty of Medicine, Research Associate, 医学部, 助手 (60351798)
Project Period (FY) 2005 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2005: ¥2,500,000 (Direct Cost: ¥2,500,000)
Keywordsamyotrophic lateral sclerosis / superoxide dismutase / Cu / Zn-SOD / monoclonal antibody / oxidation / conformational change / 脳神経疾患 / 銅代謝 / モノクローナル抗体
Research Abstract

Amyotrophic lateral sclerosis (ALS) is a neurological disease characterized by selective motor neurons in the brain and spinal cord. Although more than 100 mutations have been identified in the Copper/Zinc-superoxide dismutase (Cu/Zn-SOD) in Familial amyotrophic lateral sclerosis (FALS), the mechanism responsible for FALS remains unclear. The finding of the present study shows that FALS causing-mutant Cu/Zn-SOD proteins (FALS mutant SODs), but not wild-type SOD, are barely detected by three monoclonal antibodies (mAbs) in Western blot analyses. ELISA for denatured FALS mutant SODs by DTT, SDS or heat treatment also showed a lowered immunoreactivity against the mAbs compared with wild-type SOD. Because all the epitopes of these mAbs are mapped within the Greek key loop (residues 102-115 in human Cu/Zn-SOD), these data uggest that different conformational changes occur in the loop between wild-type and FALS mutant SODs during the unfolding process. Circular dichroism measurements revealed that FALS mutant SODs are sensitive to denaturation by DTT, SDS or heat treatment, but these results partly but not completely explain the injurious properties of FALS mutant SODs. The findings reported herein suggest that, in addition to the instability of FALS mutant SODs, subtle conformational differences in the Greek key loop between wild-type and FALS mutant SODs during/after denaturation are involved in the etiology of FALS. In addition, recent studies suggest that oxidative damage of Cu/Zn-SOD itself has own pathogenicity in ALS. Through mass spectrometry and limited proteolysis, it was determined that the mass size of the molecule is 32 and 48, and that the modification site in Cu/Zn-SOD is Cys 111 in the Greek key loop. We demonstrated that the Cys111 is selectively oxidized to cysteine sulfinic acid (Cys-S0_2H) and to cysteine sulfonic acid (Cys-SO_3H) under mild oxidizing conditions.

Report

(3 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • Research Products

    (22 results)

All 2006 2005

All Journal Article (21 results) Book (1 results)

  • [Journal Article] Effects of copper metabolism on neurological functions in Wistar and Wilson's disease model rats.2006

    • Author(s)
      Fujiwara N.
    • Journal Title

      Biochem. Biophys. Res. Commun. 349・3

      Pages: 1079-1086

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Reactive oxygen species modify oligosaccharides of glycoproteins in vivo : A study of a spontaneous acute hepatitis model rat (LEC rat).2006

    • Author(s)
      Yasuda J.
    • Journal Title

      Biochem. Biophys. Res. Commun. 342・1

      Pages: 127-134

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Inhibition of gene expression of heparin-binding epidermal growth factor-like growth factor by extracellular superoxide dismutase in rat aortic smooth muscle cells.2006

    • Author(s)
      Nishimura M.
    • Journal Title

      Free Radic. Res. 40・6

      Pages: 589-595

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Effects of copper metabolism on neurological functions in Wistar and Wilson's disease model rats.2006

    • Author(s)
      Fujiwara, N.
    • Journal Title

      Biochem Biophys Res Coommun. 349. 3

      Pages: 1079-1086

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Reactive oxygen species modify oligosaccharides of glycoproteins in vivo : a study of a spontaneous acute hepatitis model rat (LEC rat).2006

    • Author(s)
      Yasuda, J.
    • Journal Title

      Biochem Biophys Res Coommun. 342. 1

      Pages: 127-134

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Inhibition of gene expression of heparin-binding epidermal growth factor-like growth factor by extracellular superoxide dismutase in rat smooth muscle cells2006

    • Author(s)
      Nishimura, M.
    • Journal Title

      Free Radic Res. 40. 6

      Pages: 589-595

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Effects of copper metabolism on neurological functions in Wistar and Wilson's disease model rats.2006

    • Author(s)
      Fujiwara N.
    • Journal Title

      Biochem.Biophys.Res.Commun. 349・3

      Pages: 1079-1086

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Reactive oxygen species modify oligosaccharides of glycoproteins in vivo : A study of a spontaneous acute hepatitis model rat (LEC rat).2006

    • Author(s)
      Yasuda J.
    • Journal Title

      Biochem.Biophys.Res.Commun. 342・1

      Pages: 127-134

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Inhibition of gene expression of heparin-binding epidermal growth factor-like growth factor by extracellular superoxide dismutase in rat aortic smooth muscle cells.2006

    • Author(s)
      Nishimura M.
    • Journal Title

      Free Radic.Res. 40・6

      Pages: 589-595

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Different immunoreactivity against monoclonal antibodies between wild-type and mutant copper/zinc superoxide dismutase linked to amyotrophic lateral sclerosis.2005

    • Author(s)
      Fujiwara, N.
    • Journal Title

      J. Biol. Chem. 28・6

      Pages: 5061-5070

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Overexpression of Mutated Cu,Zn-SOD in Neuroblastoma Cells Results in Cytoskeletal Change.2005

    • Author(s)
      Takamiya, R.
    • Journal Title

      Am J Physiol Cell Physiol 288・2

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Modification of oligosaccharides by reactive oxygen species decreases sialyl lewis x-mediated cell adhesion.2005

    • Author(s)
      Eguchi, H.
    • Journal Title

      Glycobiology 15・11

      Pages: 1094-1101

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Different immunoreactivity against monoclonal antibodies between wild-type and mutant copper/zinc superoxide dismutase linked to amyotrophic lateral sclerosis.2005

    • Author(s)
      Fujiwara, N.
    • Journal Title

      J Biol Chem. 280.6

      Pages: 5061-70

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Overexpression of mutated Cu, Zn-SOD in neuroblastoma cells results in cytoskeletal change.2005

    • Author(s)
      Takamiya, R.
    • Journal Title

      Am J Physiol Cell Physiol 288.2

      Pages: 253-259

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Modification of oligosaccharides by reactive oxygen species decreases sialyl lewis x mediated cell adhesion.2005

    • Author(s)
      Eguchi, H.
    • Journal Title

      Glycobiology 15.11

      Pages: 1094-101

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Induction of thioredoxin reductase as an adaptive response to acrolein in human umbilical vein endothelial cells.2005

    • Author(s)
      Park YS
    • Journal Title

      Biochem. Biophys. Res. Commun. 327.4

      Pages: 1058-1065

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Clostridium botulinum type A progenitor toxin binds to intestine-407 cells via N-acetyllactosamine moiety.2005

    • Author(s)
      Kojima S
    • Journal Title

      Biochem. Biophys. Res. Commun. 331.2

      Pages: 571-576

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Tumor necrosisi factor-a-induced iron sequestration and oxidative stress in human endothelial cells.2005

    • Author(s)
      Nanami, M.
    • Journal Title

      Arterioscler Thromb Vasc Biol. 25.12

      Pages: 2495-501

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Different immunoreactivity against monoclonal antibodies between wild-type and mutant copper/zinc superoxide dismutase linked to amyotrophic lateral sclerosis.2005

    • Author(s)
      Fujiwara, N.
    • Journal Title

      J Biol Chem. 28・6

      Pages: 5061-5070

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Overexpression of Mutated Cu, Zn-SOD in Neuroblastoma Cells Results in Cytoskeletal Change.2005

    • Author(s)
      Takamiya, R.
    • Journal Title

      Am J Physiol Cell Physiol 288・2

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Modification of oligosaccharides by reactive oxygen species decreases sialyl lewis x-mediated cell adhesion.2005

    • Author(s)
      Eguchi H
    • Journal Title

      Glycobiology 15・11

      Pages: 1094-1101

    • Related Report
      2005 Annual Research Report
  • [Book] Control of NO Production. Nitric Oxide Donors for Pharmaceutical and Biological Applications2005

    • Author(s)
      Fujiwara, N.
    • Total Pages
      27
    • Publisher
      WILEY-VCH, Weinheim
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary 2005 Annual Research Report

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Published: 2005-04-01   Modified: 2016-04-21  

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