2003 年度実績報告書

DNAポリメラーゼβおよびDNAトポイソメラーゼII新規特異的阻害物質の研究

研究課題

研究課題/領域番号	14780466
研究機関	神戸学院大学
研究代表者	水品善之神戸学院大学, 栄養学部, 講師 (20307705)
キーワード	DNAポリメラーゼβ / DNAトポイソメラーゼII / 酵素阻害剤 / 蛋白質立体構造 / 抗癌剤 / コンピュータ・シミュレーション
研究概要	(1)天然材料からDNAポリメラーゼおよびDNAトポイソメラーゼII阻害物質の探索天然からDNAポリメラーゼβおよびDNAトポイソメラーゼIIを選択的に阻害する物質を探索した。薬用植物であるナンテンの葉から細胞を脱分化させて、大量培養した細胞の脂溶性抽出液に両酵素の阻害活性が見られたので、阻害物質を単離・精製した。化学構造を^1H-および^<13>C-NMR,FAB-MSで解析した結果、トリテルペンであった。そこで、この細胞に存在する各種トリテルペン類を単離・精製、化学構造を決定してから、トリテルペンの構造とDNAポリメラーゼβおよびDNAトポイソメラーゼIIの阻害活性の相関を調査した。カルボキシル基(COOH基)を有すること、水酸基(OH基)が少ないことが両酵素の阻害に必須であることをが示唆されたDNAポリメラーゼβとDNAトポイソメラーゼIIの共通阻害にはトリテルペンのステロール骨格の構造が関わっており、olide型やlupane型よりも、ursane型が阻害に重要であることがわかった。 (2)酵素-阻害剤の分子間相互作用の解析 DNAトポイソメラーゼII阻害剤として報告されているHoppospongic acid A (HA-A)がDNAポリメラーゼβを阻害することを見出した。DNAポリメラーゼβ (39kDa)はトリプシンで加水分解してdNTP結合活性を持つ31kDaドメイン断片とDNA結合活性を持つ8kDaドメイン断片に分けることができる。これら酵素や断片を用いてHAAとの分子間相互作用測定を実施した。その結果、HA-Aは8kDaドメインのssDNA結合活性に必須なアミノ酸群と結合することが解明された。そして、そのアミノ酸群を鋳型にしてDNAトポイソメラーゼIIのssDNA結合活性領域におけるHA-A結合シミュレーションに成功した。

研究成果

(25件)

すべてその他

すべて文献書誌 (25件)

[文献書誌] Yoshiyuki Mizushina et al.: "Cyanogenic glycoside from Taraxacum offcinale"Natural Med.. 57・1. 36 (2003)
[文献書誌] Yoshiyuki Mizushina et al.: "Kohamaic acid A, a novel sesterterpenic acid, inhibits activities of DNA polymerases from deuterostomes"Biochim.Biophys.Acta. 1648・1-2. 55-61 (2003)
[文献書誌] Yoshiyuki Mizushina et al.: "Molecular action mode of hippospogic acid A, an inhibitor of gastrulation of starfish embryos"J.Biochem.. 133・4. 541-552 (2003)
[文献書誌] Yoshiyuki Mizushina et al.: "The inhibitory action of pyrrolidine alkaloid, 1,4-dideoxy-1,4-imino-D-ribitol, on eukaryotic DNA polymerases"Biochem.Biophys.Res.Commun.. 304・1. 78-85 (2003)
[文献書誌] Kiminori Matsubara et al.: "Inhibitory effect of pyridoxal 5'-phosphate on endothelial cell proliferation, replicative DNA polymerase and DNA topoisomerase"Int.J.Mol.Med.. 12・1. 51-55 (2003)
[文献書誌] Chikako Murakami et al.: "Analysis of cell cycle regulation by l-mono-0-acyl-3-0-(α-D-sulfoquinovosyl)-glyceride (SQMG),an inhibitor of eukaryotic DNA polymerases"Biochem.Pharmacol.. 66・4. 541-550 (2003)
[文献書誌] Hiromi Yoshida et al.: "Variation in the content of tocopherols and istribution of fatty acids within soya bean seeds (Glycine max L.)"J.Food Comp.Anal.. 16. 429-440 (2003)
[文献書誌] Yoshiyuki Mizushina et al.: "Inhibition of DNA polymerases and DNA topoisomerase II by triterpenes produced by plant callus"Biochem.Biophys.Res.Commun.. 305・2. 365-373 (2003)
[文献書誌] Yoshiyuki Mizushina et al.: "Selective inhibition of mammalian DNA polymerase alpha by vitamin D2 and D3"J.Pharmacol.Sci.. 92・3. 283-290 (2003)
[文献書誌] Hiromi Yoshida et al.: "Effects of microwave treatment on the oxidative stability of peanut (Arachis hypogaea) oils and the molecular species of their triacylglycerols"Eur.J.Lipid.Sci.Technol.. 105. 351-358 (2003)
[文献書誌] Hiromi Yoshida et al.: "Roasting effects on the distribution of tocopherols and phospholipids within each structural part and section of soybeans"J.Am.Oil Chem.Soc.. 80・7. 665-674 (2003)
[文献書誌] Yoshiyuki Mizushina et al.: "Some anti-chronic inflammatory compounds are DNA polymerase λ-specific inhibitors"Biochem.Pharmacol.. 66・10. 1935-1944 (2003)
[文献書誌] Akira Iida et al.: "Phenylethanoid glycosides targeting mammalian DNA polymerases"Natural Med.. 57・4. 146-149 (2003)
[文献書誌] Yoshiyuki Mizushina et al.: "Inhibitory action of emulsified sulfoquinovosyl acylglycerol on mammalian DNA polymerases"Lipids. 38・10. 1065-1074 (2003)
[文献書誌] Yoshiyuki Mizushina et al.: "Pyridoxal 5'-phosphate is a selective inhibitor in vivo of DNA polymerase α and ε"Biochem.Biophys.Res.Commun.. 312・4. 1025-1032 (2003)
[文献書誌] Shim-ichi Matsuura et al.: "Novel immobilization technique of single-stranded DNA molecules on a glass surface for direct analysis of DNA polymerase activity"ISBC2003. 2003. 322-325 (2003)
[文献書誌] Kiminori Matsubara et al.: "Petasiphenol which inhibits DNA polymerase λ activity is an inhibitor of in vitro angiogenesis"Oncol.Rep.. 11・2. 447-451 (2004)
[文献書誌] Toshihiro Akihisa et al.: "Dehydrotrametenonic Acid and Dehydroeburiconic Acid from Poria cocos and Their Inhibitory Effects on Eukaryotic DNA Polymerase α and β"Biosci.Biotechnol.Biochem.. 68・2. 448-450 (2004)
[文献書誌] Isoko Kuriyama et al.: "L-Homoserylaminoethanol, a novel dipeptide alcohol inhibitor of eukaryotic DNA polymerase ε from a plant cultured cells, Nicotina tabacum L."Bioorg.Med.Chem.. 12・5. 957-962 (2004)
[文献書誌] Hiromi Yoshida et al.: "Lipid classes, fatty acid composition and triacylglycerol molecular species in the kernels of pumpkin seeds (Cucurbita spp.)"J.Sci.Food.Agric.. 84. 158-163 (2004)
[文献書誌] Chikako Murakami et al.: "Mechanism of cell cycle arrest by sulfoquinovosyl monoacylglycerol with a C18-saturated fatty acid (C18-SQMG)"Biochem.Pharmacol.. 67・7. 1373-1380 (2004)
[文献書誌] Hiromi Yoshida et al.: "Variations in the composition of acyl lipids and triacylglycerol molecular species of pumpkin (Cucurbita spp.) seeds"Eur.J.Lipid Sci.Technol.. 106. 101-109 (2004)
[文献書誌] Kouji Kuramochi et al.: "Structure-activity relationships of epolactaene analogs as DNA polymerase inhibitors"Bioorg.Med.Chem.. (in press). (2004)
[文献書誌] Hiromi Yoshida et al.: "Fatty acid distributions of triacylglycerols and phospholipids in peanuts seeds (Arachis hypogaea L.) following microwave treatment"J.Food.Comp.Anal.. (in press). (2004)
[文献書誌] Yoshiyuki Mizushina et al.: "Selective inhibition of the activities of both DNA polymerases and DNA topoisomerase II by lanostane-type triterpene acids from Poria cocos"Cancer Sci.. (in press). (2004)

2003 年度 実績報告書

DNAポリメラーゼβおよびDNAトポイソメラーゼII新規特異的阻害物質の研究

研究代表者

水品 善之 神戸学院大学, 栄養学部, 講師 (20307705)

研究成果

[文献書誌] Yoshiyuki Mizushina et al.: "Cyanogenic glycoside from Taraxacum offcinale"Natural Med.. 57・1. 36 (2003)

[文献書誌] Yoshiyuki Mizushina et al.: "Kohamaic acid A, a novel sesterterpenic acid, inhibits activities of DNA polymerases from deuterostomes"Biochim.Biophys.Acta. 1648・1-2. 55-61 (2003)

[文献書誌] Yoshiyuki Mizushina et al.: "Molecular action mode of hippospogic acid A, an inhibitor of gastrulation of starfish embryos"J.Biochem.. 133・4. 541-552 (2003)

[文献書誌] Yoshiyuki Mizushina et al.: "The inhibitory action of pyrrolidine alkaloid, 1,4-dideoxy-1,4-imino-D-ribitol, on eukaryotic DNA polymerases"Biochem.Biophys.Res.Commun.. 304・1. 78-85 (2003)

[文献書誌] Kiminori Matsubara et al.: "Inhibitory effect of pyridoxal 5'-phosphate on endothelial cell proliferation, replicative DNA polymerase and DNA topoisomerase"Int.J.Mol.Med.. 12・1. 51-55 (2003)

[文献書誌] Chikako Murakami et al.: "Analysis of cell cycle regulation by l-mono-0-acyl-3-0-(α-D-sulfoquinovosyl)-glyceride (SQMG),an inhibitor of eukaryotic DNA polymerases"Biochem.Pharmacol.. 66・4. 541-550 (2003)

[文献書誌] Hiromi Yoshida et al.: "Variation in the content of tocopherols and istribution of fatty acids within soya bean seeds (Glycine max L.)"J.Food Comp.Anal.. 16. 429-440 (2003)

[文献書誌] Yoshiyuki Mizushina et al.: "Inhibition of DNA polymerases and DNA topoisomerase II by triterpenes produced by plant callus"Biochem.Biophys.Res.Commun.. 305・2. 365-373 (2003)

[文献書誌] Yoshiyuki Mizushina et al.: "Selective inhibition of mammalian DNA polymerase alpha by vitamin D2 and D3"J.Pharmacol.Sci.. 92・3. 283-290 (2003)

[文献書誌] Hiromi Yoshida et al.: "Effects of microwave treatment on the oxidative stability of peanut (Arachis hypogaea) oils and the molecular species of their triacylglycerols"Eur.J.Lipid.Sci.Technol.. 105. 351-358 (2003)

[文献書誌] Hiromi Yoshida et al.: "Roasting effects on the distribution of tocopherols and phospholipids within each structural part and section of soybeans"J.Am.Oil Chem.Soc.. 80・7. 665-674 (2003)

[文献書誌] Yoshiyuki Mizushina et al.: "Some anti-chronic inflammatory compounds are DNA polymerase λ-specific inhibitors"Biochem.Pharmacol.. 66・10. 1935-1944 (2003)

[文献書誌] Akira Iida et al.: "Phenylethanoid glycosides targeting mammalian DNA polymerases"Natural Med.. 57・4. 146-149 (2003)

[文献書誌] Yoshiyuki Mizushina et al.: "Inhibitory action of emulsified sulfoquinovosyl acylglycerol on mammalian DNA polymerases"Lipids. 38・10. 1065-1074 (2003)

[文献書誌] Yoshiyuki Mizushina et al.: "Pyridoxal 5'-phosphate is a selective inhibitor in vivo of DNA polymerase α and ε"Biochem.Biophys.Res.Commun.. 312・4. 1025-1032 (2003)

[文献書誌] Shim-ichi Matsuura et al.: "Novel immobilization technique of single-stranded DNA molecules on a glass surface for direct analysis of DNA polymerase activity"ISBC2003. 2003. 322-325 (2003)

[文献書誌] Kiminori Matsubara et al.: "Petasiphenol which inhibits DNA polymerase λ activity is an inhibitor of in vitro angiogenesis"Oncol.Rep.. 11・2. 447-451 (2004)

[文献書誌] Toshihiro Akihisa et al.: "Dehydrotrametenonic Acid and Dehydroeburiconic Acid from Poria cocos and Their Inhibitory Effects on Eukaryotic DNA Polymerase α and β"Biosci.Biotechnol.Biochem.. 68・2. 448-450 (2004)

[文献書誌] Isoko Kuriyama et al.: "L-Homoserylaminoethanol, a novel dipeptide alcohol inhibitor of eukaryotic DNA polymerase ε from a plant cultured cells, Nicotina tabacum L."Bioorg.Med.Chem.. 12・5. 957-962 (2004)

[文献書誌] Hiromi Yoshida et al.: "Lipid classes, fatty acid composition and triacylglycerol molecular species in the kernels of pumpkin seeds (Cucurbita spp.)"J.Sci.Food.Agric.. 84. 158-163 (2004)

[文献書誌] Chikako Murakami et al.: "Mechanism of cell cycle arrest by sulfoquinovosyl monoacylglycerol with a C18-saturated fatty acid (C18-SQMG)"Biochem.Pharmacol.. 67・7. 1373-1380 (2004)

[文献書誌] Hiromi Yoshida et al.: "Variations in the composition of acyl lipids and triacylglycerol molecular species of pumpkin (Cucurbita spp.) seeds"Eur.J.Lipid Sci.Technol.. 106. 101-109 (2004)

[文献書誌] Kouji Kuramochi et al.: "Structure-activity relationships of epolactaene analogs as DNA polymerase inhibitors"Bioorg.Med.Chem.. (in press). (2004)

[文献書誌] Hiromi Yoshida et al.: "Fatty acid distributions of triacylglycerols and phospholipids in peanuts seeds (Arachis hypogaea L.) following microwave treatment"J.Food.Comp.Anal.. (in press). (2004)

[文献書誌] Yoshiyuki Mizushina et al.: "Selective inhibition of the activities of both DNA polymerases and DNA topoisomerase II by lanostane-type triterpene acids from Poria cocos"Cancer Sci.. (in press). (2004)

2003 年度実績報告書

水品善之神戸学院大学, 栄養学部, 講師 (20307705)