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著作: [鈴木 登志子]/Fujii Yutaka/Miyano Masashi/Chen Lan-Ying/Takahashi Tomohiro/Watanabe Kikuko/cDNA cloning, expression, and mutagenesis study of liver-type prostaglandin F synthase./[The Journal of Biological Chemistry]

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EID
97653
EOID
729153
Map
0
LastModified
2014年4月3日(木) 15:50:48
Operator
松井 栄里
Avail
TRUE
Censor
0
Owner
[学科長]/[徳島大学.医学部.医学科]
Read
継承
Write
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Delete
継承
種別 必須 学術論文(審査論文)
言語 必須 英語
招待 推奨
審査 推奨 Peer Review
カテゴリ 推奨
共著種別 推奨
学究種別 推奨
組織 推奨
  1. 徳島大学.大学院ヘルスバイオサイエンス研究部.神経情報医学部門.情報統合医学講座(2004年4月1日〜)
著者 必須
  1. 鈴木 登志子(->個人[山本 登志子])
    役割 任意
    貢献度 任意
    学籍番号 推奨
  2. (英) Fujii Yutaka
    役割 任意
    貢献度 任意
    学籍番号 推奨
  3. (英) Miyano Masashi
    役割 任意
    貢献度 任意
    学籍番号 推奨
  4. (英) Chen Lan-Ying
    役割 任意
    貢献度 任意
    学籍番号 推奨
  5. (英) Takahashi Tomohiro
    役割 任意
    貢献度 任意
    学籍番号 推奨
  6. (英) Watanabe Kikuko
    役割 任意
    貢献度 任意
    学籍番号 推奨
題名 必須

(英) cDNA cloning, expression, and mutagenesis study of liver-type prostaglandin F synthase.

副題 任意
要約 任意

(英) Prostaglandin (PG) F synthase catalyzes the reduction of PGD2 to 9alpha,11beta-PGF2 and that of PGH2 to PGF2alpha on the same molecule. PGF synthase has at least two isoforms, the lung-type enzyme (Km value of 120 microM for PGD2 (Watanabe, K., Yoshida, R., Shimizu, T., and Hayaishi, O. (1985) J. Biol. Chem. 260, 7035-7041) and the liver-type one (Km value of 10 microM for PGD2 (Chen, L. -Y., Watanabe, K., and Hayaishi, O. (1992) Arch. Biochem. Biophys. 296, 17-26)). The liver-type enzyme was presently found to consist of a 969-base pair open reading frame coding for a 323-amino acid polypeptide with a Mr of 36,742. Sequence analysis indicated that the bovine liver PGF synthase had 87, 79, 77, and 76% identity with the bovine lung PGF synthase and human liver dihydrodiol dehydrogenase (DD) isozymes DD1, DD2, and DD4, respectively. Moreover, the amino acid sequence of the liver-type PGF synthase was identical with that of bovine liver DD3. The liver-type PGF synthase was expressed in COS-7 cells, and its recombinant enzyme had almost the same properties as the native enzyme. Furthermore, to investigate the nature of catalysis and/or substrate binding of PGF synthase, we constructed and characterized various mutant enzymes as follows: R27E, R91Q, H170C, R223L, K225S, S301R, and N306Y. Although the reductase activities toward PGH2 and phenanthrenequinone (PQ) of almost all mutants were not inactivated, the Km values of R27E, R91Q, H170C, R223L, and N306Y for PGD2 were increased from 15 to 110, 145, 75, 180, and 100 microM, respectively, indicating that Arg27, Arg91, His170, Arg223, and Asn306 are essential to give a low Km value for PGD2 of the liver-type PGF synthase and that these amino acid residues serve in the binding of PGD2. Moreover, the R223L mutant among these seven mutants especially has a profound effect on kcat for PGD2 reduction. The Km values of R223L, K225S, and S301R for PQ were about 2-10-fold lower than the wild-type value, indicating that the amino acid residues at 223, 225 and 301 serve in the binding of PQ to the enzyme. On the other hand, the Km value of H170C for PGH2 was 8-fold lower than that of the wild type, indicating that the amino acid residue at 170 is related to the binding of PGH2 to the enzyme and that Cys170 confer high affinity for PGH2. Additionally, the 5-fold increase in kcat/Km value of the N306Y mutant for PGH2 compared with the wild-type value suggests that the amino acid at 306 plays an important role in catalytic efficiency for PGH2.

キーワード 推奨
  1. (英) Amino Acid Sequence
  2. (英) Animals
  3. (英) Base Sequence
  4. (英) Blotting, Northern
  5. (英) COS Cells
  6. (英) Cattle
  7. (英) Chromatography, Gel
  8. (英) Cloning, Molecular
  9. (英) DNA, Complementary
  10. (英) Humans
  11. (英) Hydroxyprostaglandin Dehydrogenases
  12. (英) Liver
  13. (英) Molecular Sequence Data
  14. (英) Mutagenesis, Site-Directed
  15. (英) RNA, Messenger
  16. (英) Recombinant Proteins
  17. (英) Sequence Homology, Amino Acid
  18. (英) Substrate Specificity
発行所 推奨
誌名 必須 The Journal of Biological Chemistry([The American Society for Biochemistry and Molecular Biology])
(pISSN: 0021-9258, eISSN: 1083-351X)
ISSN 任意 0021-9258
ISSN: 0021-9258 (pISSN: 0021-9258, eISSN: 1083-351X)
Title: The Journal of biological chemistry
Title(ISO): J Biol Chem
Publisher: American Society for Biochemistry and Molecular Biology
 (NLM Catalog  (Scopus  (CrossRef (Scopus information is found. [need login])
必須 274
必須 1
必須 241 248
都市 任意
年月日 必須 1999年 1月 1日
URL 任意
DOI 任意 10.1074/jbc.274.1.241    (→Scopusで検索)
PMID 任意 9867836    (→Scopusで検索)
NAID 任意
WOS 任意 000077900200036
Scopus 任意 2-s2.0-0032957688
評価値 任意
被引用数 任意
指導教員 推奨
備考 任意
  1. (英) Article.DataBankList.DataBank.DataBankName: GENBANK

  2. (英) Article.DataBankList.DataBank.AccessionNumberList.AccessionNumber: D88749

  3. (英) Article.PublicationTypeList.PublicationType: Journal Article

  4. (英) Article.PublicationTypeList.PublicationType: Research Support, Non-U.S. Gov't