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1999 Fiscal Year Final Research Report Summary

BOMBYX CYSTEINE PROTEINASE (BCP) : X-RAY DIFFRACTION ANALYSIS.

Research Project

Project/Area Number 10044210
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field 動物生理・代謝
Research InstitutionYamaguchi University

Principal Investigator

TAKAHASHI Susumu  Faculty of Agriculture, Yamaguchi University, Professor, 農学部, 教授 (90022665)

Co-Investigator(Kenkyū-buntansha) KAGEYAMA Takashi  Primate Research Institute, Kyoto University, Professor, 霊長類研究所, 教授 (20027501)
YAMAMOTO Yoshimi  Faculty of Agriculture, Yamaguchi University, Associate Professor, 農学部, 助教授 (40115514)
WATANABE Syoji  Faculty of Agriculture, Yamaguchi University, Associate Professor, 農学部, 助教授 (30113020)
Project Period (FY) 1998 – 1999
KeywordsCysteine Protease / BCP / Activation / Structure / Bombyx
Research Abstract

In the silkmoth eggs, a large amount of inactive pro-cysteine protease accumulates. BCP(47kDa) is processed in vitro into an enzymatically active 39-kDa molecule(M-form). Similar active enzyme is also found in developing silkmoth eggs, indicating that the activation of this enzyme is a key step in yolk protein degradation. In order to know the detailed mechanism of activation, we examined the structure of proBCP using X-ray crystallography, circular dichroiam and high voltage electron microscopy, as well as kinetic experiments. Gel filtration and SDS-PAGE analysis suggest that proBCP exist as an octameric structure. In this study we directly demonstrated the octameric structure of proBCP by high voltage electron microscopy. Brief exposure of proBCP to acidic conditions, it was cornverted to a monomeric structure. If it is exposed to acidic conditions for a longer period, limited proteolysis occurs at N-terminal region and converted to 39-kDa active form. Kinetic experiments suggest that the reaction might be an autocatalytic intramolecular reaction, in which monomeric pro-BCP is converted to an active enzyme through intermediate forms, X & Y, releasing small peptides. This is the major processing in the early stage of pro-BCP activation. In addition, we crysalized pro-BCP and, to confirm above theory. Furthermore, we found a novel cysteine protease specific inhibitors, cloned cDNA and amino acid sequense of it was determined.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] YAMAMOTO,Y.,Watanabe,S YAMHAMA,Y.,TAKAHASHI,S.Y.: "Bombyx acid cysteine protease(BCP):Hormonal regulation of biosynthesis and accumulation in the overy"J.INSECT PHYIOL.. 46. 783-791 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] YAMAMOTO,Y.,WATANABE,S.KAGEYAMA,T.TAKAHASHI,S.: "A novel inhibitor for bombyx systeine proteinase is homologous to propeptide reguons of cysteine proteinases"FEBS,Lett.. 448. 257-260 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] YAMAMOTO,Y.他: "Punification and charactenzation of Bombyx cycteine proteinase(BCP) specfic inhibitors from hemolymph of Bombyx morl"ARCH.INSECT BIOCHEM.PHYSIOL.. 41. 119-129 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] YAMAMOTO,Y.他: "Proregion of Bomyx mori cysteine proteinase functions as an intramolecular chaperone to promote proper folding of the mature enzyme"ARCH.INSECT BIOCHEM.PHYSIOL.. 42. 167-178 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] WATANABE,S.他: "Mitochondrial thioredoxin reductase in bovine adrena cortex.Its purification,properties,nuleotide/amino acid aequences,and identification of selenocysteine"EUR.J.BIOCHEM.. 264. 74-84 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] INOKUMA,H.他: "Duration of antibodies against 24 kDa protein of Rhipicephalus sanguineus extract dogs infested with adult ticks"J.vet.MED.SCI.. 61. 179-181 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] YAMAMOTO, Y., YAMAHAMA, Y., WATABE, S. and TAKAHASHI, S. Y.: "Bombyx acid cysteine protease (BCP) : Hormonalregulation of biosynthesis and accumulation in the ovary."J. Insect Physiol.. 46. 783-791 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] YAMAMOTO, Y., WATANABE, S., KAGEYAMA, T. and TAKAHASHI, S. Y.: "A novel inhibitor for Bombyx cysteine proteinase is homologous to propeptide regions of cysteine proteinases."FEBS, Lett.. 448. 257-260 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] YAMAMOTO, Y., WATANABE, S., KAGEYAMA, T. & TAKAHASHI, S. Y.: "Purification and characterization of Bombyx cycteine proteinase(BCP) specific inhibitors from the hemolymph of Bombyx mori."Arch. Insect Biochem.. Physio. 41. 119-129 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] YAMAMOTO, Y., WATANABE, S., KAGEYAMA, T. and TAKAHASHI, S. Y.: "Proregion of Bombyx mori cysteine proteinase functions as an intra-molecular chaperone to promote proper folding of the mature enzyme.."Arch. Insect Biochem. Physiol.. 42. 167-178 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] WATANABE, S., MAKINO, Y., OGAWA, K., HIROI, T., YAMAMOTO, Y. and TAKAHASHI, S. Y.: "Mitochondrial thioredoxin reductase in bovine adrenal cortex. Its purification, properties, nucleotide/amino acid sequences, and identification of selenocysteine."Eur. J. Biochem.. 264. 74-84 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] INOKURA, H., NUKAI, M., OHNO, K., YAMAMOTO, Y., TAKAHASHI, S. Y. & OHNISHI, T.: "Duration of antibodies against 24 kDa protein of Rhipicephalus sanguineus extract in dogs infested with adult ticks."J. vet. Med. Sci.. 61. 179-181 (1999)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2001-10-23  

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