Two-dimensional alanine scanning mutational analysis of the interaction between the vitamin D receptor and its ligands: studies of A-ring modified 19-norvitamin D analogs

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• 作者：Masato Shimizu ; Keiko Yamamoto ; Mika Mihori ; Yukiko Iwasaki ; Daisuke Morizono ; Sachiko Yamada
• 关键词：Alanine scanning mutation ; Vitamin D receptor ; 19-Norvitamin D analogs ; Structure– ; function relationship ; Chemical synthesis
• 刊名：The Journal of Steroid Biochemistry and Molecular Biology
• 出版年：2004
• 出版时间：May 2004
• 年：2004
• 卷：89-90
• 期：Complete
• 页码：75-81
• 全文大小：356 K

文摘

To clarify the structure–function relationship (SFR) of vitamin D analogs in terms of their interaction with the vitamin D receptor (VDR), we have proposed a new approach, two-dimensional alanine scanning mutational analysis (2D-ASMA). In this paper, attention was focused on the interactions around the A-ring of vitamin D. For this purpose, we synthesized four new 2-substituted 19-norvitamin D derivatives (3–6). The VDR affinity (3–6: 1, 5, 2 and 1/140, respectively) and transcriptional activity (3–6: 10, 30, 2 and 0.3, respectively) of the four compounds were evaluated relative to 1,25-(OH)₂D₃ (5) (normalized to 1). Then, the transcriptional activities of wild-type and 18 mutant VDRs induced by the four compounds (3–6) were investigated. The results of this 18×4 2D-ASMA were presented as a patch table, and the effects of the mutations were analyzed in comparison with the natural hormone (1) and 2-methylene-19-nor-20-epi-1,25-(OH)₂D₃ (2MD, 2). Of the four A-ring analogs, the 2α-hydroxyethoxy derivative (3) showed striking differences in the pattern on the patch table. From the results, we suggest a docking mode of this compound (3) in which the A-ring adopts the α conformation.