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1992 Fiscal Year Final Research Report Summary

IDENTIFICATION OF NEW SPECIES OF BRAIN-SPECIFIC CYTOCHROME b_5

Research Project

Project/Area Number 03670128
Research Category

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

Allocation TypeSingle-year Grants
Research Field General medical chemistry
Research InstitutionOITA MEDICAL UNIVERSITY

Principal Investigator

YUBISUI Toshitsugu  OITA MEDICAL UNIVERSITY, DEPARTMENT OF BIOCHEMISTRY, ASSOCIATE PROFESSOR, 医学部, 助教授 (00019564)

Co-Investigator(Kenkyū-buntansha) SHIRABE Komei  OITA MEDICAL UNIVERSITY, DEPARTMENT OF BIOCHEMISTRY, ASSISTANT PROFESSOR, 医学部, 助手 (50179058)
TAKESHITA Masazumi  OITA MEDICAL UNIVERSITY, DEPARTMENT OF BIOCHEMISTRY, PROFESSOR, 医学部, 教授 (50019551)
Project Period (FY) 1991 – 1992
KeywordsCYTOCHROME b_5 / cDNA CLONING / FUNCTIONS IN BRAIN / LOCALIZATION IN BRAIN
Research Abstract

In this research project we tried to identify new species of brain-specific cytochrome b_5 from human and mouse brain cDNA library. By plaque hybridization method we screened total of 5x10^5 plaques of mouse brain cDNA library using ECL 3'-oligolabelled primer and random-labelled cDNA of human liver. We obtained 16 clones hybridized with 3'-oligolabelled primers, and 18 clones with random-labelled cDNA. Among them 4 clones gave common signals with both probes, and other clones were all different. Isolation of insert DNAs for analyzing the base sequences of these clones are now proceeding.
In parallel with the screening the cDNA for new species for brain-specific cytochrome b_5 we analyzed the distribution of cytochrome b_5 in rat brain immunohistochemically using anti-cytochrome b_5. Cytochrome b_5 was found to localize to both white and gray matters, but especially to neuronal cells in gray matter. This is a sharp contrast to the distribution of cytochrome P-450 in brain which was shown to localize selectively to myelinated fibers in white matter. Such a clear difference in the distributions of cytochrome P-450 and cytochrome b_5 in brain tissues observed in this study is the first example.
These results apparently indicate that it is important to identify the structure of and to analyze the functions of brain-specific cytochrome b_5s. Some species of brain-specific cytochrome b_5 may deeply participate in the function of neuronal cells in gray matter.

  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] Nagai Takushi: "Analysis of Mutant NADH-Cytochrome b_5 Reductase:Apparent TypeIII Methemoglobinemia Can Be Explained as TypeI with an Unstable Reductase" Blood. 81. 808-814 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shirabe Komei: "Enzymatic Instability of NADH-cytochrome b_5 Reductase as a Cause of Hereditary Methemoglobinemia TypeI(Red Cell Type)" Journal of Biological Chemistry. 267. 20416-20421 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yubisui Toshitsugu: "Structural Role of Serine 127 in the NADH-binding Site of Human NADH-Cytochrome b_5 Reductase" Juurnal of Biological Chemistry. 266. 66-70 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Katube Takanori: "Exonic Point Mutations in NADH-Cytochrome b_5 Reductase Genes of Heterozygotes for Hereditary Methemoglobinemia TypeI and III" American Journal of Human Genetics. 48. 799-808 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shirabe Komei: "Role of Cysteine Residues in Human NADH-Cytochrome b_5 Reductase Studied by Site-directed Mutagenesis" Journal of Biological Chemistry. 266. 7531-7536 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yubisui Toshitsugu: "Studies on the Role of Serine-127 in NADH-Cytochrome b_5 Reductase by Site-directed Mutagenesis" Flavins and Fravoproteins,Proceedings of the 10th International Symposium. 856-858 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yubisui, T., Shirabe, K., Takeshita, M., Kobayashi, Y., Fukumaki, Y., Sakaki, Y. and Takano, T.: "Structural Role of Serine 127 in the NADH-binding Site of Human NADH-Cytochrome b_5 Reductase" J. Biol. Chem. 266. 66-70 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shirabe, K., Yubisui, T., Nishino, T. and Takeshita, M.: "Role of Cysteine Residues in Human NADH-Cytochrome b_5 Reductase Studied by Site-directed Mutagenesis" J. Biol. Chem. 266. 7531-7536 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Katsube, T., Sakamoto, N., Kobayashi, Y., Seki, R., Hirano, M., Tanishima, K., Tomoda, A., Takazakura, E., Yubisui, T., Takeshita, M., Sakaki, Y. and Fukumaki, Y.: "Exonic Point Mutations in NADH-Cytochrome b_5 Reductase Genes of Homozygotes for Hereditary Methemoglobinemia, Type I and III : Putative Mechanisms of Tissue-dependent Enzyme Deficiency" Am. J. Hum. Genet.48. 799-808 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takeshita, M., Ogata, K. and Yoshida, S.: "Activation of Fatty Acid in Rat Brain Microsomes" Neurochem. Res. 16. 174 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshida, S., Miyata, M., Yoshida, H. and Takeshita, M.: "Change of Conformational Disorder in Membrane Lipids in the Pulmonary Artery of Monocrotaline-injected Rats Detected in situ by Fourier Transform Infrared Spectroscopy" Vibrational Spectrosc.3. 271-276 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takeshita, M., Yoshida, S., Miyata, M. and Yoshida, H.: "Analysis of Factors in Hypertension : Change of Membrane Fluidity in the Arteries of Spontaneously Hypertensive and Monocrotaline-injected Rats" Clin. Experim. Pharmacol. Physiol.19. 33-35 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shirabe, K., Yubisui, T., Borgese, N., Tang, C. Y., Hultquist, D. E. and Takeshita, M.: "Enzymatic Instability of NADH-Cytochrome b_5 Reductase as a Cause of" J. Biol. Chem.267. 20416-20421 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ogata, K., Tamura, M. and Takeshita, M.: "Spermine Down-regulates Superoxide Generation Induced by fMet-Leu-Phe in Electropermeabilized Human Neutrophils" Biochem. Biophys. Res. Commun.182. 20-26 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nagai, T., Shirabe, K., Yubisui, T. and Takeshita, M.: "Analysis of Mutant NADH-Cytochrome b_5 Reductase : Apparent Type III Methemoglobinemia can be Explained as Type I With An Unstable Reductase" Blood. 81 (3). 808-814 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Fujimoto, Y., Shirabe, K., Nagai, T., Yubisui, T. and Takeshita, M.: "Role of Lys-110 of Human NADH-Cytochrome b_5 Reductase in NADH Binding as Probed by Site-directed Mutagenesis" FEBS Letters. (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshida, S., Yubisui, T., Shirabe K. and Takeshita, M.: "Analysis by Fourier Transform Infrared and Circular Dichroism Spectroscopies of Protein Structures of Soluble NADH-Cytochrome b_5 Reductases Prepared by Site-directed Mutagenesis- Comparison with Ferredoxin-NADP^+ Reductase-" Biochemistry. (1993)

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

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Published: 1994-03-24  

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