2002 Fiscal Year Annual Research Report

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

Research Project

Project/Area Number	14780466
Research Institution	Kobe Gakuin University
Principal Investigator	水品善之神戸学院大学, 栄養学部, 講師 (20307705)
Keywords	DNAポリメラーゼβ / DNAトポイソメラーゼII / 酵素阻害剤 / タンパク質立体構造 / 抗癌剤 / コンピュータ・シミュレーション
Research Abstract	(1)天然材料からのDNAポリメラーゼβおよびDNAトポイソメラーゼII阻害物質の探索これまでに見いだした長鎖不飽和脂肪酸(ネルボン酸)およびある種のステロイド(リトコール酸)の他に、新規なDNAポリメラーゼβおよびDNAトポイソメラーゼIIを特異的に阻害する物質を探索した。両酵素の阻害活性を指標にして、インド産の海綿動物(Plakinastrella sp.)から阻害物質を単離・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、Elenic acidであった。本物質はDNAポリメラーゼβおよびDNAトポイソメラーゼIIを選択的に阻害して、ヒト胃癌細胞の増殖を抑制した[Mizushina et al.,Biochem. Pharmacol.(2002)63,399-407]。新規の抗癌剤になることが期待される。 (2)酵素-阻害剤の分子間相互作用の解析 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阻害物質(ネルボン酸およびリトコール酸)との分子間相互作用測定を実施した。その結果、ネルボン酸とリトコール酸は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阻害物質結合シミュレーションに成功した[水品他,蛋白質核酸酵素(2002)47,708-714]。ネルボン酸はHIV逆転写酵素も阻害することから、同様にssDNA結合活性領域にけるドッキング・シミュレーションを行った[Kasai, Mizushina et al., J. Biochem.(2002)132,819-828]。

Research Products

(23 results)

All Other

All Publications (23 results)

[Publications] Yoshiyuki Mizushina et al.: "A plant phytotoxin, solanapyrone A, is an inhibitor of DNA polymerase β and λ"J. Biol. Chem.. 277・1. 630-638 (2002)
[Publications] Hiroeki Sahara et al.: "Anti-tumor effect of chemically synthesized sulfolipids based on sea urchin's natural sulfonoquinovosylmonoacylglycerols"Jpn. J. Cancer Res.. 93・1. 85-92 (2002)
[Publications] Madoka Watanabe et al.: "Biotinylated lithocholic acids for affinity chromatography of mammalian DNA polymerases α and β"Bioorg. Med. Chem. Lett.. 12・3. 287-290 (2002)
[Publications] Yoshiyuki Mizushina et al.: "Selective inhibition of the activities of both eukaryotic DNA polymerases and DNA topoisomerases by elenic acid"Biochem. Pharmacol.. 63・3. 399-407 (2002)
[Publications] Shinji Kamisuki et al.: "Actions of derivatives of dehydroaltenusin, a new mammalian DNA polymerase α-specific inhibitor"Biochem. Pharmacol.. 63・3. 421-427 (2002)
[Publications] Chikako Murakami et al.: "Vitamin A-related compounds, all-trans retinal and retinoic acids, selectively inhibit activities of mammalian replicative DNA polymerases"Biochim. Biophys. Acta. 1574・1. 85-92 (2002)
[Publications] Chikako Murakami et al.: "Novel anti-inflammatory compounds from Rubus sieboldli, terpenoids, are inhibitors of mammalian DNA polymerases"Biochim. Biophys. Acta. 1596・2. 193-200 (2002)
[Publications] Chikako Murakami et al.: "Structure-function relationship of synthetic sulfoquinovosyl-acylglycerols as a mammalian DNA polymerases inhibitors"Arch. Biochem. Biophys.. 403・2. 229-236 (2002)
[Publications] Yoshitaka Matsumoto et al.: "An immunosuppressive effect by synthetic sulfonolipids deduced from sulfonoquinovosyl diacylglycerols of sea urchin"Transplantation. 74・2. 261-267 (2002)
[Publications] Masako Oda et al.: "Inhibition of telomerase by linear-chain fatty acids : A structural analysis"Biochem. J.. 367・2. 329-334 (2002)
[Publications] Nobuyuki Kasai et al.: "Three-dimensional structural model analysis of binding site of an inhibitor, nervonic acid, of both DNA polymerase β and HIV-1 reverse transcriptase"J. Biochem.. 132・5. 819-828 (2002)
[Publications] Yoshiyuki Mizushina et al.: "Petasiphenol : a DNA polymerase λ inhibitor"Biochemistry. 41・49. 14463-14471 (2002)
[Publications] 坂口謙吾, 菅原二三男, 水品善之: "真核生物DNAポリメラーゼの阻害剤:ケミカルノックアウト剤およびサプレッサー様天然物"生化学(日本生化学会). 47・3. 29-32 (2002)
[Publications] 水品善之他: "分子プローブを用いたDNAポリメラーゼβとDNAトポイソメラーゼIIの立体構造相関の解析"蛋白質核酸酵素(共立出版). 47・6. 708-714 (2002)
[Publications] Chikako Murakami et al.: "Effects of glycolipids from spinach on mammalian DNA polymerases"Biochem. Pharmacol.. 65・2. 259-267 (2003)
[Publications] Chikako Murakami et al.: "A novel DNA polyrnerase inhibitor and a potent apoptosis inducer : 2-mono-0-acy1-3-0-(α-D-sulfoquinovosyl)-glyceride with one stearic acid"Biochim. Biophys. Acta. 1645・1. 72-80 (2003)
[Publications] Yoshiyuki Mizushina et al.: "Flavonoid glycoside : a new inhibitor of eukaryotic DNA polymerase α, and a new carrier for inhibitor-affinity chromatography"Biochem. Biophys. Res. Commun.. 301・2. 480-487 (2003)
[Publications] Shunya Takahashi et al.: "Total synthesis of dehydroaltenusin"Tetrahedron Lett.. 44・9. 1875-1877 (2003)
[Publications] Yoshiyuki Mizushina et al.: "A sulfoquinovosyl diacylglycerol is a DNA polymerase ε inhibitor"Biochem. J.. 370・1. 299-305 (2003)
[Publications] Yoshiyuki Mizushina et al.: "Kohamaic acid A, a novel sesterterpenic acid, inhibits activities of DNA polymerases from deuterostomes"Biochim. Biophys. Acta. (in press). (2003)
[Publications] Yoshiyuki Mizushina et al.: "Cyanogenic glycoside from Taraxacum officinale"Natural Med.. (in press). (2003)
[Publications] Yoshiyuki Mizushina et al.: "Molecular action mode of hippospogic acid A, an inhibitor of gastrulation of starfish embryos"J. Biochem.. (in press). (2003)
[Publications] Chikako Murakami et al.: "Analysis of cell cycle regulation by 1-mono-0-acy1-3-0-(α-D-sulfoquinovosyl)-glyceride (SQMG), an inhibitor of eukaryotic DNA polymerases"Biochem. Pharmacol.. (in press). (2003)

2002 Fiscal Year Annual Research Report

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

Principal Investigator

水品 善之 神戸学院大学, 栄養学部, 講師 (20307705)

Research Products

[Publications] Yoshiyuki Mizushina et al.: "A plant phytotoxin, solanapyrone A, is an inhibitor of DNA polymerase β and λ"J. Biol. Chem.. 277・1. 630-638 (2002)

[Publications] Hiroeki Sahara et al.: "Anti-tumor effect of chemically synthesized sulfolipids based on sea urchin's natural sulfonoquinovosylmonoacylglycerols"Jpn. J. Cancer Res.. 93・1. 85-92 (2002)

[Publications] Madoka Watanabe et al.: "Biotinylated lithocholic acids for affinity chromatography of mammalian DNA polymerases α and β"Bioorg. Med. Chem. Lett.. 12・3. 287-290 (2002)

[Publications] Yoshiyuki Mizushina et al.: "Selective inhibition of the activities of both eukaryotic DNA polymerases and DNA topoisomerases by elenic acid"Biochem. Pharmacol.. 63・3. 399-407 (2002)

[Publications] Shinji Kamisuki et al.: "Actions of derivatives of dehydroaltenusin, a new mammalian DNA polymerase α-specific inhibitor"Biochem. Pharmacol.. 63・3. 421-427 (2002)

[Publications] Chikako Murakami et al.: "Vitamin A-related compounds, all-trans retinal and retinoic acids, selectively inhibit activities of mammalian replicative DNA polymerases"Biochim. Biophys. Acta. 1574・1. 85-92 (2002)

[Publications] Chikako Murakami et al.: "Novel anti-inflammatory compounds from Rubus sieboldli, terpenoids, are inhibitors of mammalian DNA polymerases"Biochim. Biophys. Acta. 1596・2. 193-200 (2002)

[Publications] Chikako Murakami et al.: "Structure-function relationship of synthetic sulfoquinovosyl-acylglycerols as a mammalian DNA polymerases inhibitors"Arch. Biochem. Biophys.. 403・2. 229-236 (2002)

[Publications] Yoshitaka Matsumoto et al.: "An immunosuppressive effect by synthetic sulfonolipids deduced from sulfonoquinovosyl diacylglycerols of sea urchin"Transplantation. 74・2. 261-267 (2002)

[Publications] Masako Oda et al.: "Inhibition of telomerase by linear-chain fatty acids : A structural analysis"Biochem. J.. 367・2. 329-334 (2002)

[Publications] Nobuyuki Kasai et al.: "Three-dimensional structural model analysis of binding site of an inhibitor, nervonic acid, of both DNA polymerase β and HIV-1 reverse transcriptase"J. Biochem.. 132・5. 819-828 (2002)

[Publications] Yoshiyuki Mizushina et al.: "Petasiphenol : a DNA polymerase λ inhibitor"Biochemistry. 41・49. 14463-14471 (2002)

[Publications] 坂口謙吾, 菅原二三男, 水品善之: "真核生物DNAポリメラーゼの阻害剤:ケミカルノックアウト剤およびサプレッサー様天然物"生化学(日本生化学会). 47・3. 29-32 (2002)

[Publications] 水品善之 他: "分子プローブを用いたDNAポリメラーゼβとDNAトポイソメラーゼIIの立体構造相関の解析"蛋白質 核酸 酵素(共立出版). 47・6. 708-714 (2002)

[Publications] Chikako Murakami et al.: "Effects of glycolipids from spinach on mammalian DNA polymerases"Biochem. Pharmacol.. 65・2. 259-267 (2003)

[Publications] Chikako Murakami et al.: "A novel DNA polyrnerase inhibitor and a potent apoptosis inducer : 2-mono-0-acy1-3-0-(α-D-sulfoquinovosyl)-glyceride with one stearic acid"Biochim. Biophys. Acta. 1645・1. 72-80 (2003)

[Publications] Yoshiyuki Mizushina et al.: "Flavonoid glycoside : a new inhibitor of eukaryotic DNA polymerase α, and a new carrier for inhibitor-affinity chromatography"Biochem. Biophys. Res. Commun.. 301・2. 480-487 (2003)

[Publications] Shunya Takahashi et al.: "Total synthesis of dehydroaltenusin"Tetrahedron Lett.. 44・9. 1875-1877 (2003)

[Publications] Yoshiyuki Mizushina et al.: "A sulfoquinovosyl diacylglycerol is a DNA polymerase ε inhibitor"Biochem. J.. 370・1. 299-305 (2003)

[Publications] Yoshiyuki Mizushina et al.: "Kohamaic acid A, a novel sesterterpenic acid, inhibits activities of DNA polymerases from deuterostomes"Biochim. Biophys. Acta. (in press). (2003)

[Publications] Yoshiyuki Mizushina et al.: "Cyanogenic glycoside from Taraxacum officinale"Natural Med.. (in press). (2003)

[Publications] Yoshiyuki Mizushina et al.: "Molecular action mode of hippospogic acid A, an inhibitor of gastrulation of starfish embryos"J. Biochem.. (in press). (2003)

[Publications] Chikako Murakami et al.: "Analysis of cell cycle regulation by 1-mono-0-acy1-3-0-(α-D-sulfoquinovosyl)-glyceride (SQMG), an inhibitor of eukaryotic DNA polymerases"Biochem. Pharmacol.. (in press). (2003)

水品善之神戸学院大学, 栄養学部, 講師 (20307705)

[Publications] 水品善之他: "分子プローブを用いたDNAポリメラーゼβとDNAトポイソメラーゼIIの立体構造相関の解析"蛋白質核酸酵素(共立出版). 47・6. 708-714 (2002)