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Structural biological analysis of activation mechanism of hematocyte by ENF peptide family

Research Project

Project/Area Number 16370049
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

KAWANO Keiichi  Hokkaido Univ., Fac.Sci., Prof., 大学院理学研究院, 教授 (10136492)

Co-Investigator(Kenkyū-buntansha) HAYAKAWA Yoichi  Saga Univ., Fac.Agric., Prof., 農学部, 教授 (50164926)
KAWABATA Shun-ichiro  Kyushu Univ., Fac.Sci., Prof., 大学院理学研究科, 教授 (90183037)
MIZUGUCHI Mineyuki  Univ.Toyama., Fac.Pharm., Assoc.Prof., 大学院医学薬学研究部, 助教授 (30332662)
AIZAWA Tomoyasu  Hokkaido Univ., Fac.Adv.Life Sci., Assoc.Prof., 大学院先端生命科学研究院, 助教授 (40333596)
DEMURA Makoto  Hokkaido Univ., Fac.of Adv.Life Sci., Prof., 大学院先端生命科学研究院, 教授 (70188704)
Project Period (FY) 2004 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥14,500,000 (Direct Cost: ¥14,500,000)
Fiscal Year 2006: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2005: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2004: ¥7,900,000 (Direct Cost: ¥7,900,000)
Keywordsinnate immune system / protein / conformational analysis / peptide / invertebrate / GBP / サイトカイン / 細胞成長因子 / 昆虫 / 生体防御 / 立体構造
Research Abstract

Growth-blocking peptide (GBP) is a small insect cytokine with various activities: larval growth regulation, cell proliferation, and stimulation of a class of lepidopteran insect blood cells (plasmatocytes). GBP was initially identified as a 25 amino acid peptide hormone (1-25 GBP), which suppress the growth of host armyworm, found in the hemolymph of the parasitized armyworm. However, in non-parasitized armyworm larvae, 23 amino acid GBP is expressed in insect plasma. Subsequent studies revealed that the parasitization by wasp induced an elongation of the C-terminal amino acid sequence of GBP (1-28 GBP). In this study, we characterized the GBP analogs, which consist of various length of C-terminal region, by comparison of their biological activities and three-dimensional structures. The results indicate that 1-28 GBP exhibits the strongest activity to suppress the larval growth. NMR analysis shows that these peptides have the same tertiary structures basically, ^-which consist of a dis … More ordered N-terminal region, a structured core stabilized by a disulfide bond and an antiparallel β-sheet in aqueous solution. However, the C-terminal region of 1-28 GBP undergoes conformational transition from a random-coiled state to an a-helical state in the presence of dodecylphosphocholine (DPC) micelles. It suggests that the C-terminal region bound to membrane would have an affects on the larval growth activity.
We isolated, identified, and determined the solution structure of a novel peptide from lepidopteran, Pseudaletia separata that induces rapid hemocyte aggregation. The isolated peptide from crude integument extract, designated hemocyte-aggregation factor (AGF), was revealed to contain 32 amino acid residues (H_2N-SVQILRCPDGMQMLRSGQCVATTEPPFDPDSY-COOH) by N-terminal amino acid sequencing and 3'-RACE. NMR spectra of ^<15>N-labelled AGF were recorded at 283-313 K and pH 5.0. The full sequential assignment for all non-proline residues was achieved using ^<15>N-edited TOCSY and NOESY spectra. The solution structure was calculated by CYANA and discussed in relevance to the functional feature of AGF. Less

Report

(4 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • 2004 Annual Research Report
  • Research Products

    (44 results)

All 2007 2006 2005 2004

All Journal Article (43 results) Book (1 results)

  • [Journal Article] Equilibrium and kinetics of the folding and unfolding of canine milk lysozyme.2007

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

      Biochemistry (in press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] The solution structure of horseshoe crab antimicrobial peptide tachystatin B with an inhibitory cystine-knot motif.2007

    • Author(s)
      Fujitani, N
    • Journal Title

      J. Peptide Sci 13(4)

      Pages: 269-279

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Structural approach to a novel tandem repeat DNA-binding domain, STPR, by CD and NMR.2007

    • Author(s)
      Saito, S
    • Journal Title

      Biochemistry 46(7)

      Pages: 1703-1713

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Destabilization of transthyretin by pathogenic mutations in the DE loop2007

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

      Proteins 66(3)

      Pages: 716-725

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] The solution structure of horseshoe crab antimicrobial peptide tachystatin B with an inhibitory cystine-knot motif.2007

    • Author(s)
      Fujitani, N
    • Journal Title

      J.Peptide Sci. 13(4)

      Pages: 269-279

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Structural approach to a novel tandem repeat DNA-binding domain, STPR, by CD and NMR.2007

    • Author(s)
      Saito, S.
    • Journal Title

      Biochemistry 46(7)

      Pages: 1703-1713

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Destabilization of transthyretin by pathogenic mutations in the DE loop.2007

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

      Proteins, 66(3)

      Pages: 716-725

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] The solution structure of horseshoe crab antimicrobial peptide tachystatin B with the inhibitory cystine-knot motif2007

    • Author(s)
      N.Fujitani
    • Journal Title

      J. Peptide Science (in press)

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Preparation and observation of fresh-frozen sections of the green fluorescent protein transgenic mouse head.2006

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

      Acta Histochemica et Cytochemica 39(2)

      Pages: 31-34

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] N-Terminal mutational analysis of the interaction between growth-blocking peptide (GBP) and receptor of insect immune cells.2006

    • Author(s)
      Watanabe, S.
    • Journal Title

      Protein & Peptide Letters 13(8)

      Pages: 815-822

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] A novel N14Y mutation in connexin26 in keratitis-ichthyosis-deafness syndrome. Analyses of altered gap junctional communication and molecular structure of N terminus of mutated connexin26.2006

    • Author(s)
      Arita, K.
    • Journal Title

      Am. J. Pathol 169(2)

      Pages: 416-423

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] A novel N14Y mutation in connexin26 in keratitis-ichthyosis-deafness syndrome. Analyses of altered gap junctional communication and molecular structure of N terminus of mutated connexin26.2006

    • Author(s)
      Arita, K.
    • Journal Title

      Am.J.Pathol 169(2)

      Pages: 416-423

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Preparation and observation of fresh-frozen sections of the green fluorescent protein transgenic mouse head.2006

    • Author(s)
      M.Tada
    • Journal Title

      Acta Histochemica et Cytochemica 39(2)

      Pages: 31-34

    • Related Report
      2006 Annual Research Report
  • [Journal Article] A novel N14Y mutation in connexin26 in keratitis-ichthyosis-deafness syndrome. -Analyses of altered gap junctional communication and molecular structure of N terminus of mutated connexin26-2006

    • Author(s)
      K.Arita
    • Journal Title

      Am. J. Pathol 169(2)

      Pages: 416-423

    • Related Report
      2006 Annual Research Report
  • [Journal Article] N-Terminal mutational analysis of the interaction between growth-blocking peptide (GBP) and receptor of insect immune cells2006

    • Author(s)
      S.Watanabe
    • Journal Title

      Protein & Peptide Letters 13(8)

      Pages: 815-822

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Solution structure of microtubule-associated protein light chain 3 and identification of its functional subdomains.2005

    • Author(s)
      Kouno, T.
    • Journal Title

      J. Biol. Chem. 280(26)

      Pages: 24610-24617

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Dimeric transthyretin variant assembles into spherical neurotoxins.2005

    • Author(s)
      Matsubara, K.
    • Journal Title

      Biochemistry 44(9)

      Pages: 3280-3288

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Structure of the central hub of bacteriophage Mu baseplate determined by X-ray crystallography of gp442005

    • Author(s)
      Kondou, Y.
    • Journal Title

      J. Mol. Biol. 352(4)

      Pages: 976-985

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Disassembling and bleaching of chloride-free pharaonis halorhodopsin by octyl-β-glucoside.2005

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

      Biochemistry 44(39)

      Pages: 12923-12931

    • 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 the stabilization of the molten globule state on the folding mechanism of α-lactalbumin : A study of a chimera of bovine and human α-lactalbumin.2005

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

      Proteins 61(2)

      Pages: 356-365

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Peptide mimics of epidermal growth factor (EGF) with antagonistic activity.2005

    • Author(s)
      Nakamura, T.
    • Journal Title

      J. Biotechnology 116(3)

      Pages: 211-219

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Crystallization and preliminary X-ray analysis of gene product 44 from bacteriophage Mu.2005

    • Author(s)
      Kondou, Y.
    • Journal Title

      Acta Cryst. F 61

      Pages: 104-105

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Solution structure of microtubule- associated protein light chain 3 and identification of its functional subdomains.2005

    • Author(s)
      Kouno, T.
    • Journal Title

      J.Biol.Chem. 280(26)

      Pages: 24610-24617

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Structure of the central hub of bacteriophage Mu baseplate determined by X-ray crystallography of gp44.2005

    • Author(s)
      Kondou, Y.
    • Journal Title

      J.Mol.Biol. 352(4)

      Pages: 976-985

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Peptide mimics of epidermal growth factor (EGF) with antagonistic activity.2005

    • Author(s)
      Nakamura, T.
    • Journal Title

      J.Biotechnology 116(3)

      Pages: 211-219

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Crystallization and preliminary X-ray analysis of gene product 44 from bacteriophage Mu.2005

    • Author(s)
      Kondou, Y.
    • Journal Title

      Acta Cryst.F. 61

      Pages: 104-105

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Peptide mimics of epidermal growth factor(EGF)with antagonistic activity.2005

    • Author(s)
      Nakamura, T.
    • Journal Title

      J.Biotechnology 116

      Pages: 211-219

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Ab initio MO computation of the hydration effect on the ionization potential of sodium pyrenesulfonate.2005

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

      J.Mol.Struct.(THEOCHEM) 715

      Pages: 205-214

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Dimeric transthyretin variant assembles into spherical neurotoxins.2005

    • Author(s)
      Matsubara, K.
    • Journal Title

      Biochemistry 44

      Pages: 3280-3288

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Crystallization and preliminary X-ray analysis of gene product 44 from bacteriophage Mu.2005

    • Author(s)
      Kondou, Y.
    • Journal Title

      Acta Cryst.F 61

      Pages: 104-105

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Effects of the stabilization of the molten globule state on the folding mechanism of α-lactalbumin: A study of a chimera of bovine and human α-lactalbumin.2005

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

      Proteins 61

      Pages: 356-365

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Solution structure of microtubule-associated protein light chain 3 and identification of its functional subdomains.2005

    • Author(s)
      Kouno, T.
    • Journal Title

      J.Biol.Chem 280

      Pages: 24610-24617

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Disassembling and bleaching of chloride-free pharaonis, halorhodopsin by octyl-β-glucoside.2005

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

      Biochemistry 44

      Pages: 12923-12931

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Structure of the central hub of bacteriophage Mu baseplate determined by X-ray crystallography of gp44.2005

    • Author(s)
      Kondou, Y.
    • Journal Title

      J.Mol.Biol. 352

      Pages: 976-985

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Peptide mimics of epidermal growth factor (EGF) with antagonistic activity.2005

    • Author(s)
      Nakamura, T. 他
    • Journal Title

      J.Biotechnology 116

      Pages: 211-219

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Dimeric transthyretin variant assembles into spherical neurotoxins.2005

    • Author(s)
      Matsubara, K. 他
    • Journal Title

      Biochemistry 44

      Pages: 3280-3288

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Ab initio MO computation of the hydration effect on the ionization potential of sodium pyrenesulfonate.2005

    • Author(s)
      Shinoda, H. 他
    • Journal Title

      J.Mol.Struct.(THEOCHEM) 715

      Pages: 205-214

    • Related Report
      2004 Annual Research Report
  • [Journal Article] ^1H, ^<13>C and ^<15>N resonance assignments of human microtubule-associated protein light chain-32004

    • Author(s)
      Kouno, T.
    • Journal Title

      J. Biomol. NMR 29(3)

      Pages: 415-416

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] The Gly-Gly linker region of the insect cytokine growth blocking peptide (GBP) is essential for activity.2004

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

      J. Biol. Chem. 279(49)

      Pages: 51331-51337

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] ^1H, ^<13>C and ^<15>N resonance assignments of human microtubule-associated protein light chain-3.2004

    • Author(s)
      Kouno, T.
    • Journal Title

      J.Biomol.NMR 29(3)

      Pages: 415-416

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] The Gly-Gly linker region of the insect cytokine growth blocking peptide (GBP) is essential for activity.2004

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

      J.Biol.Chem. 279(49)

      Pages: 51331-51337

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] The Gly-Gly linker region of the insect cytokine growth blocking peptide (GBP) is essential for activity.2004

    • Author(s)
      Yoshida, M. 他
    • Journal Title

      J.Biol.Chem. 279

      Pages: 51331-51337

    • Related Report
      2004 Annual Research Report
  • [Journal Article] ^1H, ^<13>C and ^<15>N resonance assignments of human microtubule-associated protein light chain-3.2004

    • Author(s)
      Kouno, T. 他
    • Journal Title

      J.Biomol.NMR 29

      Pages: 415-416

    • Related Report
      2004 Annual Research Report
  • [Book] ゲノミクス・プロテオミクスの新展開 (今中忠行監修、共同執筆)2004

    • Author(s)
      水口峰之, 他
    • Total Pages
      1158
    • Publisher
      エヌ・ティー・エス
    • Related Report
      2004 Annual Research Report

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Published: 2004-04-01   Modified: 2016-04-21  

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