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Studies on the structure and function of aspartic proteases and their genes

Research Project

Project/Area Number 60440103
Research Category

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

Allocation TypeSingle-year Grants
Research Field 物質生物化学
Research InstitutionThe University of Tokyo

Principal Investigator

TAKAHASHI Kenji  Faculty of Science, The University of Tokyo; Professor, 理学部, 教授 (70011533)

Project Period (FY) 1985 – 1988
Project Status Completed (Fiscal Year 1988)
Budget Amount *help
¥31,900,000 (Direct Cost: ¥31,900,000)
Fiscal Year 1988: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1987: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1986: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1985: ¥25,000,000 (Direct Cost: ¥25,000,000)
KeywordsAspartic protease / Pepsinogen / Primary structure / Gene / Activation mechanism / Site-directed mutagenesis / DNA methylation / DNAメチル化 / cDNA / 塩基配列 / 活性化 / メチル化 / Rhizopusペプシン / Slowーmoving protease / プレペプシノーゲン / ペプシン / エクソン / イントロン / プロガストリックシン / カテプシンD / シグナルペプチド
Research Abstract

1. Human, porcine and rat prepepsinogens and calf preprochymosin were isolated and identified, and the complete amino acid sequences of their N-terminal signal sequences determined.
2. Various pepsinogens and related aspartic proteases were isolated and their molecular and enzymatic properties investigated, including human pepsinogens A, human pepsinogen C, human slow moving protease, monkey prostate pepsinogen C-like zymogen, and turtle, frog and tuna pepsinogens.
3. By using the protein-chemistry techniques, the complete primary structures were determined for monkey pepsinogens A and C, tuna pepsinogen, rhizopuspepsin, and Aspergillus niger Type A acid protease, and partial primary structures for human pepsinogens A and C, frog pepsinogens and Scytalidium lignicolum acid protease. In addition, the complete primary structures of human and rat pepsinogens C and turtle pepsinogen were deduced from the nucleotide sequence analyses of their corresponding DNAs.
4. The activation mechanisms of … More human pepsinogens A and C, monkey and porcine pepsinogens, and various other pepsinogens were investigated and compared in detail.
5. Various mutants of porcine pepsinogen were prepared by site-directed mutagenesis, and expressed in E. coli. The change of either one of the two active-site aspartic acids to glutamic acid abolished completely the potential pepsin activity.
6. The nucleotide sequences of all the nine exons and the 5'- and 3'-flanking regions of human and rat pepsinogens C were determined, and human pepsinogen C was located on the chromosome 6. Further, analyses of human pepsinogens A showed that there exist at least three different genes and that at least two of them are tandemly aligned on a human chromosomal DNA.
7. A positive correlation was found between the expression of rat and human pepsinogens C genes and their level of demethylation. In addition, a 25-Da transcriptional regulation factor was found in the nuclear fraction of rat gastric mucosa cells, which binds 5'-upstream region of the gene. Less

Report

(5 results)
  • 1988 Annual Research Report   Final Research Report Summary
  • 1987 Annual Research Report
  • 1986 Annual Research Report
  • 1985 Annual Research Report
  • Research Products

    (68 results)

All Other

All Publications (68 results)

  • [Publications] Y.Ichihara: The Journal of Biochemistry. 98. 483-492 (1985)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] A.Moriyama: The Journal of Biochemistry. 98. 1255-1261 (1985)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Takahashi: Aspartic Proteinases and Their Inhibitors Walter de Gruyter & Co.,Berlin.265-282 (1985)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Kageyama: Journal of Biological Chemistry. 261. 4395-4405 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Kageyama: Journal of Biological Chemistry. 261. 4406-4419 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Hayano: Biochemical and Biophysical Research Communications. 138. 289-296 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Y.Ichihara: European Journal of Biochemistry. 161. 7-12 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Takahashi: Journal of Biological Chemistry. 262. 1468-1478 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Kageyama: European Journal of Biochemistry. 165. 483-490 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Hayano: Journal of Biological Chemistry. 263. 1382-1385 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Takahashi: The Journal of Biochemistry. 103. 162-167 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M.Ichinose: Cancer Research. 48. 1603-1609 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M.Ichinose: Biochemical and Biophysical Research Communications. 151. 275-282 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] N.Tsukagoshi: Gene. 65. 285-292 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M.Ichinose: Biochemical and Biophysical Research Communications. 155. 670-677 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M.Tanji: European Journal of Biochemistry. 177. 251-259 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] O.Matsuzaki: Biomedical Research. 9. 515-523 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Kageyama: The Journal of Biochemistry. 105. 15-22 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Ishihara: Journal of Biological Biochemistry. (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Takahara: Cytogenetics and Cell Genetics. (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] S.B.P.Athauda: European Journal of Biochemistry. (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Y.Ichihara: "Isolation of Human, Swine, and Rat Prepepsinogens and Calf Preprochymosin, and Determination of the Primary Structures of Their NH_2-Terminal Signal Sequences." The Journal of Biochemistry. 98. 483-492 (1985)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] A.Moriyama: "Purification of Japanese Monkey Prostate Acid Protease Zymogen and Its Identification as a Pepsinogen C-Like Zymogen." The Journal of Biochemistry. 98. 1255-1261 (1985)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Takahashi: "Multiplicity and Intermediates of the Activation Mechanism of Zymogens of Gastric Aspartic Proteinases." Aspartic Proteinases and Their Inhibitors Walter de Gruyter & CO., Berlin.265-282 (1985)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Kageyama: "The Complete Amino Acid Sequence of Monkey Pepsinogen A." Journal of Biological Chemistry. 261. 4395-4405 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Kageyama: "The Complete Amino Acid Sequence of Monkey Progastricsin." Journal of Biological Chemistry. 261. 4406-4419 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Hayano: "Close Linkage of Human Chromosomal Pepsinogen A Genes." Biochemical and Biophysical Research Communications. 138. 289-296 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] Y.Ichihara: "Nucleotide Sequence of a Nearly Full-Length cDNA Coding for Pepsinogen of Rat Gastric Mucosa." European Journal of Biochemistry. 161. 7-12 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Takahashi: "The Amino Acid Sequence of Rhizopuspepsin, an Aspartic Proteinase from Rhizopus chinensis." Journal of Biological.Chemistry. 262. 1468-1478 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Kageyama: "Activation Mechanism of Monkey and Porcine Pepsinogens A, One-step and Stepwise Activation Pathways and Their Relation to Intramolecular and Intermolecular Reactions." European Journal of Biochemistry. 165. 483-490 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Hayano: "Primary Structure of Human Pepsinogen C Gene." Journal of Biological Chemistry. 263. 1382-1385 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Takahashi: "Determination of the Amino Acid Sequences of the Two Major Isozymes of Rhizopuspepsin." The Journal of Biochemistry. 103. 162-167 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M.Ichinose: "DNA Methylation and Expression of the Rat Pepsinogen Gene in Embryonic, Adult, and Neoplastic Tissues." Cancer Research. 48. 1603-1609 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M.Ichinose: "Hypomethylation and Expression of Pepsinogen A Genes in the Fundic Mucosa of Human Stomach." Biochemical and Biophysical Research Communications. 151. 275-282 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] N.Tsukagoshi: "Nucleotide Sequence and Expression in Escherichia coli of cDNA of Swine Pepsinogen: Involvement of the Amino-Terminal Portion of the Activation Peptide Segment in Restoration of the Functional Protein." Gene. 65. 285-292 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M.Ichinose: "Cell-Specific Hypomethylation of the Pepsinogen Gene in Pepsinogen-Producing Cells." Biochemical and Biophysical Research Communications. 155. 670-677 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M.Tanji: "Tuna Pepsinogens and Pepsins, Purification, Characterization and Amino-Terminal Sequences." European Journal of Biochemistry. 177. 251-259 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] O.Matsuzaki: "Improved Purification of Slow Moving Protease from Human Gastric Mucosa and Its Action on the B Chain of Oxidized Bovine Insulin." Biomedical Research. 9. 515-523 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Kageyama: "Difference of Activation Processes and Structure of Activation Peptides in Human Pepsinogens A and Progastricsin." The Journal of Biochemistry. 105. 15-22 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Ishihara: "Primary Structure and Transcriptional Regulation of Rat Pepsinogen C Gene." Journal of Biological Chemistry. in press. (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Takahara: "Assignment of Human Pepsinogen C Gene to Chromosome 6." Cytogenetics and Cell Genetics. in press. (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] S.B.P.Athauda: "Human Pepsinogens and Pepsins. Purification, Characterization and Some Structural Differences among Isozymogens and Isozymes." European Journal of Biochemistry. in press. (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M.Ichinose: Cancer Research. 48. 1603-1609 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] M.Ichinose: Biochemical and Biophysical Research Communications. 151. 275-282 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] N.Tsukagoshi: Gene. 65. 285-292 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] M.Ichinose: Biochemical and Biophysical Research Communications. 155. 670-677 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] M.Tanji: European Journal of Biochemistry. 177. 251-259 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] O.Matsuzaki: Biomedical Research. 9. 515-523 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] T.Kageyama: The Journal of Biochemistry. 105. 15-22 (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] T.Ishihara: Journal of Biological Chemistry. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] K.Takahara: Cytogenetics Cell Genetics. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] S.B.P.Athauda: European Journal Biochemistry. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] K. Takahashi: Journal of Biological Chemistry. 262. 1468-1478 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] T. Kageyama: European Journal of Biochemistry. 165. 483-490 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] K. Takahashi: Journal of Biochemistry. 103. 162-167 (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] T. Hayano: Journal of Biological Chemistry. 263. 1382-1385 (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] M. Ichinose: Cancer Research. 48. (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] M. Ichinose: Biochemical and Biophysical Research Communications. (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] T.Kageyama: Journal of Biological Chemistry. 261. 4395-4405 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] T.Kageyama: Journal of Biological Chemistry. 261. 4406-4419 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] T.Hayano: Biochemical and Biophysical Research Communications. 138. 289-296 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Y.Ichihara: European Journal of Biochemistry. 161. 7-12 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] K.Takahashi: Journal of Biological Chemistry. (1987)

    • Related Report
      1986 Annual Research Report
  • [Publications] T.Kageyama: European Journal of Biochemistry. (1987)

    • Related Report
      1986 Annual Research Report
  • [Publications] Journal of Biochemistry. 97-4. (1985)

    • Related Report
      1985 Annual Research Report
  • [Publications] Journal of Biochemistry. 98-2. (1985)

    • Related Report
      1985 Annual Research Report
  • [Publications] Journal of Biochemistry. 98-5. (1986)

    • Related Report
      1985 Annual Research Report
  • [Publications] 生化学. 57-8. (1985)

    • Related Report
      1985 Annual Research Report

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Published: 1987-03-31   Modified: 2016-04-21  

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