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1993 Fiscal Year Final Research Report Summary

Comparative studies on the moleular architecture of insitol-trisphosphate receptor and ryanodine receptor and their intracellular distribution

Research Project

Project/Area Number 04454123
Research Category

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

Allocation TypeSingle-year Grants
Research Field General anatomy (including Histology/Embryology)
Research InstitutionThe University of Tokyo

Principal Investigator

KATAYAMA Eisaku  Institute of Medical Science, The univ of Tokyo Ass.Professor, 医科学研究所, 助教授 (50111505)

Project Period (FY) 1992 – 1993
Keywordsinositol-trisphosphate receptor / ryanodine-receptor / quick-freeze electron microscopy / Ca^<2+>-channel / molecular architecture / computer image-analysis / three-dimentional reconstruction / Purkinje cell
Research Abstract

Both inositol-1,4,5-trisphosphate receptor (IP_3R) and ryanodine-receptor (RyaR) form ligand-gated Ca^<2+> -channel and play a crucial role in intracellular signal transduction. Though the total amino-acid sequence was already determined for both, histological studies on these receptors still remain insufficient and nothing substantial has been done on their three-dimensional distribution in cells or tissues. On the structure side, the molecular architecture of tetrameric Rya-R particle has been extensively studied either by negative staining or cryo-electron microscopy, leading to the reconstruction of its three-dimensional structure. Despite many efforts similarly done for IP_3-R,however, nobody has successfully obtained, so far, the concrete information on the molecular architecture of that receptor. The author has been applying his expertised technique of quick-freeze deep-etch replica electron microscopy to the studies of various kinds of protein architecture. With this technique, one obtain very conrasty images of macromolecules in solution or in cells/tissues with very high time-and spacial-resolution, with nicely preserved three-dimensional organization. Thus, we could succesfully obtain the images of IP_3R in situ in the dendrites of cerebellar Purkinje cells. We compared them with those of RyaR and were surprized to find that the physical size of IP_3R is by far smaller than the value calculated from the ratio of molecular weights of two receptors and the actual size of RyaR.We also found that tetrameric IP_3R forms a very regular two-dimensional molecular array in situ in the cell, under certain anoxemic conditions. Further studies must be done to understand the physiological role of such array. It is also of great importance to determine the higher-resolution structure of the receptor molecules to define the sites of interacton with variuos ligands.

  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] E.Katayama: "Native structure and arrangemet of inusitol-1,4,5-tris-phosphate receptor molecules in bovine cerebellar Purkinje cells ds studied by quick-freeze deep-etch elcctron microscopy." EMBO Journal. 15. 4844-4851 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] E.Katayama: "Geometry of the flagellar motor in the cykoplasmic membrane of Salmonella typhimurium as determined by stereo-photogram metry of quick-freeze deep-etch repicc images" Journal of Molecular Biology. 255. 458-475 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] E.P.Morris: "Evaluation of high resolution shadowing applied to freeze-fractured,deep-etched particles : 3-D helical reconstruction of shadowed actin tilaments" Journal of Structural Biology. 113. 47-55 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] E,Katayama: "Current Methods in Muscle Physiology : Advantages,Problems and Limitations" Oxford Vniversity Press, (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] E.Katayama: "Mechanisms of Work Production and Work Adsorption in Muscle" Plenum Press (in press),

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] E.Katayama et al.: "Native structure and arrangement of inositol-1,4,5-trisphasphote receptor molecules in brine cerebellar Purkinje cells as studied by quick-freeze deep-etch electron microscopy" EMBO.J.15. 4844-4851 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] E.Katayama et al.: "Geometry of the flagellar motor in the cytoplasmic membrane of selmonella typhymurium as determined by stereo-photogramtry of quick-freeze deep-etch replica images" I.Mol.Biol.255. 458-475 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] E.P.Morris et al.: "Evaluation of high resolution shadowing applied to freeze-fractured, deep-etched particles : 3D helical reconstruction of shadowed actin filaments" J.Struet.Biol.113. 47-55 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] E.Katayama: Electronmicroscopy coupled with quick-freezing, In Current Methods in Muscle Physiology : Advantages, Problems and Limitations. Oxford Univ.Press, (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] E.Katayama et al.: Three-dimensional Image Analysis of myosin head as captured by quick-freeze deep-etch electron microscopy, In Mechanism of Work Production and work absorption in muscle. Plenum Press (in Press),

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

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Published: 1999-03-16  

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