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The quantification of ratios between apo to holo types of metal binding protein : a new indicator of the oxidative stress in cells

Research Project

Project/Area Number 11672314
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory medicine
Research InstitutionOsaka Medical College

Principal Investigator

NAKANISHI Toyofumi  Osaka Medical College, Faculty of Medicine Assistant Professor, 医学部, 講師 (10247843)

Co-Investigator(Kenkyū-buntansha) MIYAZAKI Ayako  Osaka Medical College, Faculty of Medicine Research Associate, 医学部, 助手 (20298772)
KISHIKAWA Masahiko  Osaka Medical College, Faculty of Medicine Research Associate, 医学部, 助手 (40268199)
SHIMIZU Akira  Osaka Medical College, Faculty of Medicine Professor, 医学部, 教授 (00028581)
Project Period (FY) 1999 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2001: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2000: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1999: ¥2,000,000 (Direct Cost: ¥2,000,000)
KeywordsNanoESIMS method / Ration of Apo / Holo / Oxidative stress in the body / Cooper-binding protein / Active oxide species / Free cooper ion / Neurodegenerative disease / ホロ型 / 遊離銅イオン毒性
Research Abstract

We applied nano-electrospray ionization mass spectrometry (NanoESIMS) for the quantification of the rations between apo to holo types of metalloproteins (copper/zinc binding superoxide dismutase:SOD-1, metallothioneine) to clarify the etiology of neurodegenerative diseases such as familial amyotrophic lateral sclerosis. We examined the optimal conditions of ultra-sensitive mass spectrometric quantification of the ratios using metalloproteins standard and purified metalloproteins.
1) We examined the condition of pH range of sample solutions, contents of organic solvent and temperature of heated capillary.
2) We applied inductively coupled plasma mass spectrometry (ICPMS) instead of nanoESIMS technique.
Using the commercial SOD-1 standard, we could not detect dimer, holo type even though we modified some analytical conditions. We give up the construction of the optimal condition of NanoESIMS technique. On the other hand, we applied the ICPMS technique or the de e turn of holo type of SOD-1 and succeeded holo type of the metalloprotem. This technique (ICPMS) was a powerful tool of the quantification of the ratios between apo to holo types of metalloproteins.

Report

(4 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] Toyohumi Nakanishi, Ayako Miyazaki, Akira Shimizu, Akihumi Yamaguchi, Satoshi Nishimura: "Assessment of the Effect of Hemoglobin Variants on Routine HbAlc Measurements by Electrospray Ionization Mass Spectrometry"Clin. Chem. Acta. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Toyofumi Nakanishi, Reiko Koyama, Tsunehiko Ikeda, Akira Shimizu: "Catalogue of soluble proteins In the human vitreous humor : comparison between diabetic retinopathy and macular hole"J. Chromatography B. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T.Nakanishi, A.Shimizu: "Ratio of the ion peak intensity of glycated and non-glycated hexapeptides from the N-terminal of hemoglobin b-chain by LC-ESI mass spectrometry"Jpn. J. Soc. Mass Spectrom. 247・6. 389-391 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T.Nakanishi, A.Miyazaki, K.Iguchi, A.Shimizu: "Effect of hemoglobin variants on routine HbAlc measurements, assessed by a mass spectrometric method"Clin. Chem.. 46・10. 1689-1692 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T.Nakanishi, A.Shimizu: "Determination of ionization efficiency of glycated and non-glycated peptides from the N-terminal of hemoglobin beta-chain electrospnay ionization mass spectrometry"J. Chromatgr.(B). 746・1. 83-90 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] M.Kishikawa, T.Nakanishi, A.Shimizu, M.Yoshino.: "Detection by mass spectrometry of highly increased amount of S-sulfonated transthyretin in serum from a patient with molybdenum cofactor deficiency"Pcdiatr. Res.. 47・1. 492-494 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] M. Saraswathi, T.Nakanishi, A.Shimizu: "Relative quantification of glycated Cu-Zn superoxide dismutase in erythrocytes by electrospray ionization mass spectrometry"Biochim. Biophys. Acta. 1426 (2). 483-490 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] M. Kishikawa, T. Nakanishi, A. Miyazaki, A. Shimizu: "A simple and reliable methods of detecting variant transthyretins by multidimensional liquid chromatography coupled to electrospray ionization mass spectrometry. Amyloid"Int. J. Exp. Clin. Invest.. 6 (1). 48-53 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] A. Miyazaki, T. Nakaniashi, M. Kishikawa, T. Nakagawa, A. Shimizu, AHM. Mawjood, Y. Aoki, M. Kikuchi: "Compound heterozygosity for b+thalassemia -31(A→G) and a variant with low oxygen affinity, Hb Sagami [b139(H17)Asn→Thr]"Hemoglobin. 23 (3). 267-271 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] M. Kishikawa, T. Nakanishi, A. Miyazaki, A. Shimizu: "Enhanced amyloidogenicity of sulfonated transthyretin in vitro, hypothetical etiology of senile. Amyloid"Int. J. Exp. Clin. Invest.. 6 (3). 183-186 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] M. Kishikawa, T. Nakanishi, A. Miyazaki, A. Shimizu, H. Kusaka: "A new amyolidogenic transthyretin variant, [D38A], detected by electrospray ionization/mass spectrometry. Amyloid"Int. J. Exp. CIin. Invest.. 6 (4). 278-281 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T. Nakanishi, A. Shimizu: "Ratio of the ion peak intensity of glycated and non-glycated hexapeptides from the N-terminal of hemoglobin β-chain by LC-ESI mass spectrometry"Jpn.J. Soc. Mass Spectrom.. 247 (6). 389-391 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T. Nakanishi, A. Miyazaki, K. Iguchi, A. Shimizu: "Effect of hemoglobin variants on routine HbAlc measurements, assessed by a mass spectrometric method"Clin. Chem.. 46 (10). 1689-1692 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T. Nakanishi, A. Shimizu: "Determination of ionization efficiency of glycated and non-glycated peptides from the N-terminal of hemoglobin beta-chain by electrospray ionization mass spectrometry"J. Chromatgr. (B). 746 (1). 83-90 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T. Hiratsuka, M. Nakazato, T. Minematsu, N. Chiono, T. Nakanishi, A. Shimizu, M. Nakazato, S. Matsukura: "Structural analysis of human β-defensine-1 and its significance in urinary tract infection"Nephron. 85 (1). 34-40 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] M. Kishikawa, T. Nakanishi, A. Shimizu, M. Yoshino: "Detection by mass spectrometry of highly increased amount of S-sulfonated transthyretin in serum from a patient with molybdenum cofactor deficiency"Pediatr. Res.. 47. 492-494 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] A. Shimizu, T. Nakanishi, M. Kishikawa, A. Miyazaki: "Detection and identification of protein variants and adducts in blood and tissues: an application of soft ionization mass spectrometry to clinical diagnosis"J. Chromatography B. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] A. Shimizu, T. Nakanishi, A. Miyazaki: "Determination of an accurate ratio of a peptide, and its adduct by ESIMS: calculation of the weighted sum of singly and doubly charged ion intensity"Advances in Mass Spectrometry, Vol.15, Editors: E Gelpi et al., John Wiley & Sons, Ltd. , Chichester. 371-372

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] K. Fukada, S. Nagano, M. Satoh, C. Tohyama, T. Nakanishi, A. Shimizu, T. Yanagihara, S. Sakoda: "Stabilization of mutant Cu/Zn superoxide dismutase (SOD1) protein by co-expressed wild SOD1 protein causes disease acceleration in familial amyotrophic lateral sclerosis model mice"Eur. J. Neurosci.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Toyofumi Nakanishi, Ayako Miyazaki, Akira Simizu, Akifumi Yamaguchi, Satoshi Nishimura: "Assessment of the Effect of Hemoglobin Variants on Routine NbAlc Measurements by Electrospray Ionization Mass Spectrometry"Clin. Chem. Acta. (in press).

    • Related Report
      2001 Annual Research Report
  • [Publications] Toyofumi Nakanishi, Reiko Koyama, Tsunehiko Ikeda, Akira Shimizu: "Catalogue of soluble proteins In the human vitreous humor : comparison between diabetic retinopathy and macular hole"J. Chromatography B. (in press).

    • Related Report
      2001 Annual Research Report
  • [Publications] Nakanishi T,Miyazaki A,Iguchi K: "Effect of hemoglobin variants on routine HbA1c measurements, assessed by a mass spectrometric method."Clinical Chemistry. 46・10. 1689-1692 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Nakanishi T & Shimizu A.: "Determination of ionization efficiency of glycated and non-glycated peptides from the N-terminal of hemoglobin beta-chain by electrospray ionization mass spectrometry."J.Chromatography (B). 746・1. 83-90 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Hiratsuka T,Nakazato M,Minematsu T,Chino N,Nakanishi T,Shimizu A,Nakazato M & Matsukura S.: "Structual analysis of human b-defensine-1 and its significance in urinary tract infection."Nephron. 85. 34-40 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kishikawa M,Nakanishi T,Shimizu A & Yoshino M.: "Detection by mass spectrometry of highly increased amount of S-sulfonated transthyretin in serum from a patient with molybdenum"Pediatric Research. 47. 492-494 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Saraswathi M,Nakanishi T,& Shimizu A.: "Relative quantification of glycated Cu-Zn superoxide dismutase in erythrocytes by electrospray ionization mass spectrometry"Biochim.Biophys.Acta. 1426・2. 483-490 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kishikawa M,Nakanishi T,Miyazaki A & Shimizu A.: "A simple and reliable method of detecting variant transthyretins by multidimensional liquid chromatography coupled to electrospray ionization/mass spectrometry"Amyloid:Int J Exp Clin Invest. 6・1. 48-53 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Miyazaki A,Nakanishi T,Kishikawa M,Nakagawa T,Shimizu A,Mawjood AHM,Imai K,Aoki Y & Kikuchi M.: "Compound heterozygosity for b+-thalassemia-31(A->G)and a variant with low oxygan affinity,Hb Sagami[b139(H17)Asn->Thr]"Hemoglobin.. 23・3. 267-271 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kishikawa M,Nakanishi T,Miyazaki A & Shimizu A.: "Enhanced amyloidogenicity of sulfonated transthyretin in vitro,a hypothetical etiology of senile"Amyloid: Int J Exp Clin Invest. 6・3. 183-186 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Kishikawa M,Nakanishi T,Miyazaki A,Shimizu A & Kusaka H.: "A new amyloidogenic transthyretin variant,[D38A],detected by electrospray ionization/mass spectrometry"Amyloid: Int J Exp Clin Invest. 6・4. 278-281 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Nakanishi T & Shimizu A: "Ratio of the ion peak intensity of glycated and non-glycated hexapeptides from the N-terminal of hemoglobin β-chain by LC-ESI mass spectrometry"Jpn.Soc.Mas Spectrom.. 247・6. 389-391 (2000)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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