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1993 Fiscal Year Final Research Report Summary

Purification and cDNA cloning of human red cell pyrimidine 5'-nucleotidase isozymes

Research Project

Project/Area Number 04671405
Research Category

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

Allocation TypeSingle-year Grants
Research Field Human genetics
Research InstitutionOkinaka Memorial Institute for medical Research

Principal Investigator

HIRONO Akira  Okinaka Memorial Institute for medical Research, 専任研究員 (90181221)

Co-Investigator(Kenkyū-buntansha) FUJII Hisaichi  東京女子医科大学, 輸血部, 助教授 (70107762)
MIWA Shiro  (財)冲中記念成人病研究所, 所長 (40034954)
Project Period (FY) 1992 – 1993
KeywordsPyrimidine 5'-nucleotidase / Isozymes / Pyrimidine 5'-nucleotidase deficiency / Purification / Hemolytic anemia
Research Abstract

Pyrimidine 5'-uncleotidase (P5N) is important in clinical medicine because the deficiency of the enzyme causes a hereditary nonspherocytic hemolytic anemia. We have been carrying out the structural analysis of P5N isozymes for the purpose of elucidating the primary cause of P5N deficiency. For the first step, we have tried the purification of P5N-II isozyme, Using standard chromatographical techniques, ninety micro grams of P5N-II were purified about 134,000-fold from 2,000 ml of human red blood cells. The purified enzyme had a specific activity of 44,130 mU/mg and gave a main band on SDS polyacry lamide gel electrophoresis. Kinetic studies using the purified P5N-II revealed that the enzyme had a unique property of using a suger phosphate ester 6-phosphogluconate as one of its potent substrates. This might be important in elucidating the physiological function of this isozyme. Although we had performed the amino acid compsition analysis and the partial amino acid sequencing of the enzyme, the N-terminal amino acid residue was found to be blocked. We have already constructed a cDNA library from human reticulocytes, and deblocking and further sequencing of amino acid residues N-terminal region for cDNA cloning is in progress.

  • Research Products

    (9 results)

All Other

All Publications (9 results)

  • [Publications] Hirono A et al.: "Molecular abnormality of G6PD Konan and G6PD Ube,the most common glucose-6-phosphate dehydrogenase variants in Japan." Human Genetics. 91. 507-508 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hirono A et al.: "Human glucose-6-phosphate dehydrogenase:Structure and function of normal and variant enzymes." Haematologia. 25. 85-97 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hirono A et al.: "G6PD Nara:A new class 1 glucose-6-phosphate dehydrogenase variant with an eight amino acid deletion." Blood. 82. 3250-3252 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hirono A et al.: "Molecular abnormality of a unique Japanese glucose-6-phosphate dehydrogenase variant (G6PD Kobe) with an extremely..." American Journal of Hematology. 45. 185-186 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hirono A, Fujii H, Hirono K, Kannno H, Miwa S: "Molecular abnormality of a Japanese glucose-6-phosphate dehydrogenase variant(G6PD Tokyo) associated with hereditary non-spherocytic hemolytic anemia." Hum Genet. 88. 347-348 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hirono A, Fujii H, Miwa S: "Molecular abnormality of G6PD Konan and G6PD Ube, the most common glucose-6-phosphate dehydrogenase variants in Japan." Hum Genet. 91. 507-508 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hirono A, Miwa S: "Human glucose-6-phosphate dehydrogenase : structure and function of normal and variant enzymes." Haematologia. 25. 85-97 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hirono A, Fujii H, Shima M, Miwa S: "G6PD Nara : A new class I glucose-6-phophate dehydorogenase variant with an eight amino acid deletion." Blood. 82. 3250-3252 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hirono A, Nakayama S, Fujii H, Miwa S: "Molecular abnormality of a unique Japanese glucose-6-phosphate dehydrogenase variant (G6PD Kobe) with an extremely increased affinity for galactose 6-phosphate." Am ja Hematol. (in press).

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

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Published: 1995-03-27  

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