Structural Relationship of Lithocholic Acid Derivatives Binding to the N-Terminal 8-kDa Domain of DNA Polymerase
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- 作者：Yoshiyuki Mizushina ; Nobuyuki Kasai ; Katsumi Miura ; Shinya Hanashima ; Masaharu Takemura ; Hiromi Yoshida ; Fumio Sugawara ; and Kengo Sakaguchi
- 刊名：Biochemistry
- 出版年：2004
- 出版时间：August 24, 2004
- 年：2004
- 卷：43
- 期：33
- 页码：10669 - 10677
- 全文大小：364K
- 年卷期：v.43,no.33(August 24, 2004)
- ISSN：1520-4995
文摘
We reported previously that lithocholic acid (LCA, 3-
-hydroxy-5-
-cholan-24-oic acid), oneof the major compounds in the secondary bile acids, selectively inhibited the activity of mammalian DNApolymerase (pol ) [Mizushina, Y., Ohkubo, T., Sugawara, F., and Sakaguchi, K. (2000) Biochemistry39, 12606-12613]. The purpose of this study was to investigate the molecular structural relationship ofLCA and its 10 chemically synthesized derivatives. The inhibitory activities of pol by some derivativecompounds were stronger than that by LCA, and these compounds bound tightly to the 8-kDa domainfragment but not to the 31-kDa domain fragment of pol . Biacore analysis demonstrated that the 8-kDadomain bound selectively to compound 9 (3--O-lauroyl-5--cholan-24-oic acid), which was the strongestpol inhibitor tested, as a 1:1 complex with a dissociation constant (Kd) of 1.73 nM. From computermodeling analysis (i.e., molecular dynamics analysis), the 8-kDa domain had two inhibitor binding areas.Three amino acid residues (Lys60, Leu77, and Thr79) of the 8-kDa domain bound to LCA and compound2 (3--methoxy-5--cholan-24-oic acid), and four amino acid residues (Leu11, Lys35, His51, and Thr79)of the 8-kDa domain bound to compound 9. From these results, the structure-function relationship amongpol and its selective inhibitors was discussed.
-hydroxy-5--cholan-24-oic acid), oneof the major compounds in the secondary bile acids, selectively inhibited the activity of mammalian DNApolymerase (pol ) [Mizushina, Y., Ohkubo, T., Sugawara, F., and Sakaguchi, K. (2000) Biochemistry39, 12606-12613]. The purpose of this study was to investigate the molecular structural relationship ofLCA and its 10 chemically synthesized derivatives. The inhibitory activities of pol by some derivativecompounds were stronger than that by LCA, and these compounds bound tightly to the 8-kDa domainfragment but not to the 31-kDa domain fragment of pol . Biacore analysis demonstrated that the 8-kDadomain bound selectively to compound 9 (3--O-lauroyl-5--cholan-24-oic acid), which was the strongestpol inhibitor tested, as a 1:1 complex with a dissociation constant (Kd) of 1.73 nM. From computermodeling analysis (i.e., molecular dynamics analysis), the 8-kDa domain had two inhibitor binding areas.Three amino acid residues (Lys60, Leu77, and Thr79) of the 8-kDa domain bound to LCA and compound2 (3--methoxy-5--cholan-24-oic acid), and four amino acid residues (Leu11, Lys35, His51, and Thr79)of the 8-kDa domain bound to compound 9. From these results, the structure-function relationship amongpol and its selective inhibitors was discussed.