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Characterization of D-amino acid containing protein from sun-damaged skin of elderly donors.

Research Project

Project/Area Number 14390032
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 広領域
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

FUJII Noriko  Kyoto University, Research Reactor Institute, Professor, 原子炉実験所, 教授 (90199290)

Co-Investigator(Kenkyū-buntansha) TAJIMA Shingo  National Defense Medical College, Professor, 教授 (60129525)
SHIMO-OKA Tadashi  Asahi Techno Glass Corp, Life Science Center, Director, ライフサイエンスセンター長
SAITO Takeshi  Kyoto University, Research Reactor Institute, Lecturer, 原子炉実験所, 助手 (10274143)
Project Period (FY) 2002 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥10,800,000 (Direct Cost: ¥10,800,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2003: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2002: ¥8,200,000 (Direct Cost: ¥8,200,000)
KeywordsAging / UV irradiation / sun-damage / D-amino acid / Racemization / Isomerization / skin / Posttranslational modification
Research Abstract

1)Biologically uncommon D-beta-aspartic acid-containing protein in elastic fibers of sun damaged skin.
We have prepared the antibody against D-beta-Asp containing protein and examined its immunoreactivity in skin. The antibody recognized disintegrated elastic fibers in sun-exposed skin. Western plot analysis of the proteins isolated from sun-damaged skin demonstrated that the polypeptide with 50kDa was immunoreactive with the antibodies for D-beta-Asp containing protein and elastin. This result suggests that UV irradiation siginificant racemization and isomerization of Asp resideus in proteins of elastic fibers and the unusual proteins were accumulated during aging. Subsequnetly, we identified some D-beta-Asp containing proteins by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and the Mascot online database searching algorithm.
We propose that the antibody could be a useful indicator for sun damage of the skin.
2) Kinetic Study of racemization of aspartyl residues in model peptides corresponding to sections of elastin
We synthesized three Asp-containing model peptides corresponding to sections of elastin:1) GVADAAAA, 2) REGDPSSS, 3) AGADEGVR. We calculated the activation energy of the racemization of Asp residues in the three peptide and the time required for the Asp D/L ratio to approximate to 1.0 at body temperature(37℃). The racemization of Asp in the peptides progressed on the basis of the first order reaction. The Asp residue in the peptide No2 is the most susceptible to racemization, while the Asp residue in the peptide No1 is the least susceptible. However, there was little difference the racemization rate of Asp among three peptides. At 37℃, the time required for the D/L ratios of Asp in these peptides to 0.999 was calculated to be 50-100 years. These results suggested that the racemization of Asp occurred in skin of elderly humans.

Report

(4 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (26 results)

All 2004 2003 2002 Other

All Journal Article (14 results) Publications (12 results)

  • [Journal Article] Immunohistochemical study of chronological and photo-induced aging skins using the antibody raised against D-Aspartyl residue-containing peptide.2004

    • Author(s)
      Miura Y., Fujimoto N., Komatsu T., Tajima S., Kawada A., Saito T., Fujii N
    • Journal Title

      J.Cutan.Pathol. 31

      Pages: 51-51

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Homochirality and Life2004

    • Author(s)
      Fujii, N, Satio, T
    • Journal Title

      Chemical Record 4

      Pages: 267-267

    • NAID

      110001713368

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Kinetic Study of Racemization of Aspartyl Residues in Synthetic Elastin Peptides.2004

    • Author(s)
      Kuge K., Fujii N., Miura Y., Tajima S., Saito T.
    • Journal Title

      Amino Acids 27

      Pages: 193-193

    • NAID

      110003723344

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] The damaging effect of UV-C irradiation on lens α-crystallin2004

    • Author(s)
      Fujii N., Uchida H., Saito T.
    • Journal Title

      Molecular Vision 10

      Pages: 814-814

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Comparison of post-translational modifications of alpha A-crystallin from normal and hereditary cataract rats.2004

    • Author(s)
      Fujii N., Takeuchi N., Fujii N., Tezuka T., Kuge K., Takata T., Kamei A.
    • Journal Title

      Amino Acids 26

      Pages: 147-147

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Homochirality and Life2004

    • Author(s)
      Fujii, N, Satio, T.
    • Journal Title

      Chemical Record

      Pages: 267-278

    • NAID

      110001713368

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Kinetic Study of Racemization of Aspartyl Residues in Synthetic Elastin Peptides.2004

    • Author(s)
      Kuge K., Fujii N., Miura Y., Tajima.S., Saito T.
    • Journal Title

      Amino Acids 27

      Pages: 193-197

    • NAID

      110003723344

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The damaging effect of UV-C irradiation on lens α-crystallin.2004

    • Author(s)
      Fujii N., Uchida H., Saito T.
    • Journal Title

      Molecular Vision 10

      Pages: 810-814

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] Immunohistochemical study of chronological and photo-induced aging skins using the antibody raised against D-Aspartyl residue-containing peptide.2004

    • Author(s)
      Miura Y., Fujimoto N., Komatsu T., Tajima S., Kawada A., Saito T., Fujii N.
    • Journal Title

      J.Cutan.Pathol. 31

      Pages: 51-56

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Homochirality and Life2004

    • Author(s)
      Fujii, N, Saito, T.
    • Journal Title

      Chemical Record 4

      Pages: 267-278

    • NAID

      110001713368

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Comparison of post-translational modifications of alpha A-crystallin from normal and hereditary cataract rats.2004

    • Author(s)
      Fujii N., Takeuchi N., Fujii N., Tezuka T., Kuge K., Takata T., Kamei A., Saito T.
    • Journal Title

      Amino Acids 26

      Pages: 147-152

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Characterization of alpha A-crystallin from high molecular weight aggregates in the normal human lens.2003

    • Author(s)
      Fujii N., Awakura M., Takemoto L., Inomata M., Takata T., Fujii N., Saito T.
    • Journal Title

      Molecular Vision 9

      Pages: 315-322

    • Related Report
      2004 Annual Research Report
  • [Journal Article] The presence of D-β-aspartic acid-containing peptides in elastic fibers of sun-damaged skin : potent marker for ultraviolet-induced skin aging2002

    • Author(s)
      Fujii N., Tajima S., Tanaka N., Fujimoto N.Takata T., Shimo-Oka T.
    • Journal Title

      Biochemical Biophysical Research Communications 194

      Pages: 1047-1047

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The presence of D-β-aspartic acid-containing peptides in elastic fibers of sun-damaged skin : potent marker for ultraviolet-induced skin aging.2002

    • Author(s)
      Fujii N., Tajima S., Tanaka N., Fujimoto N., Takata T., Shimo-Oka T.
    • Journal Title

      Biochemical Biophysical Research Communications 194

      Pages: 1047-1052

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] 藤井紀子: "D-アミノ酸からみた老化の研究"化学と教育. 52. 10-13 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Miura Y, Fujimoto N, Komatsu T, Tajima S, Kawada A, Saito T, Fujii N.: "Immunohistochemical study of chronological and photo-induced aging skins using the antibody raised agaist C-Aspartyl residue-containing peptide"J.Cutan.Pathol. 31. 51E-56E (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] M.Fukushima, H.Tamate, N.Fujii, Y.Nakano: "Determination of metal-binding proteins in soft tissues of rock oyster by neutron activation analysis and particle induced X-ray emission"J.Radioanal.Nucl.Chem.. 257. 105-108 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Noriko Fujii, Minori Awakura, Larry Takemoto, Mitsushi Inomata, Takumi Takata, Norihiko Fujii, Takeshi Saito: "Characterization of αA-crystallin from high molecular weight aggregates in the normal human lens"Molecular Vision. 9. 315-322 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Noriko Fujii, Takumi Takata, Tadashi Shimo-Oka, Shingo Tajima, Yoshiko Tanaka, Takeshi Saito: "Immunological detection of D-β-aspartate-containing protein in lens derived cell lines. Dawei Yang"Molecular Vision. 9. 200-204 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Sadakane.Y., Yamazaki.T., Nakagomi.K., Akizawa.T., Fujii.N., Tanimura.T., Kaneda.M., Hatanaka.Y.: "Quantification of the isomerization of Asp residue in recombinant human aA-crystallin by reversed-phase HPLC"J.Pharm.Biomed.Anal.. 30. 1825-1833 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] N.Fujii, S.Tajima, N.Tanaka, N.Fujimoto, T.Takata, T.Shimo-Oka: "The presence of D-beta-aspartic acid-containing peptides in elastic fibers of sun-damaged skin : A potent marker for ultraviolet-induced skin aging"Biochem. Biophys. Res. Comm. 294. 1047-1051 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] N.Fujii: "D-amino acids in living higher organisms"Origin of Life and Evolution of the Biosphere. 2002. 103-127 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] M.Kaneda, K.Nakagomi, Y.Sadakane, T.Yamazaki, T.Tanimura, T.Akizawa, N.Fujii, Y.Hatanaka.: "A Rapid HPLC Determination of the Isomerization Level of ASP-Residues within Synthetic Alpha A-Crystallin Fragments"Journal of Liquid Chromatography & Related Technologies. 25. 2445-2454 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] K.Nakagomi, K.Takatsu, S.Takagi, H.Ebisu, Y.Sadakane, N.Fujii, T.Akezawa, T.Tanimura, Y.Hatanaka.: "Isolation of cathepsin B inhibitory peptides, Cabin-A1 and -A2, from a tryptic and chymotryptic hydrolysate of human serum albumin"Peptides. 23. 1567-1571 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] K.Nakagomi, A.Fujimura, H.Maeda, Y.Sadakane, N.Fujii, T.Akizawa, T.Tanimura, Y.hatanaka.: "Isolation of novel peptides, cabin-1,-2,-3 and -4,that inhibit cathepsin B from a thremolysin digest of human plasma"Biol Pharm Bull. 25. 564-568 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 藤井紀子: "太陽紫外線によるタンパク質損傷"太陽紫外線防御研究会. 12. 47-53 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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