Studies on response element of vitamin D target genes and discrimination of vitamin D actions

维生素D靶基因反应元件研究及维生素D作用判别

• 批准号: 14571997
• 负责人: YAMAMOTO Keiko
• 金额: $ 1.6万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571997/
• 关键词: vitamin D; vitamin D receptor; one-point mutant; vitamin D analog; transcriptional activity; VDRE; オステオポンチン; VDR

We investigated the interactions between the vitamin D receptor (VDR) and its ligands by a new method which is termed two-dimensional alanine scanning mutational analysis (2D-ASMA). We made one-point mutants to alanine of all amino acid residues lining the ligand binding pocket of the VDR and evaluated their transcriptional activity using 12 ligands by luciferase assay method. The results were presented on two-dimensional table with each mutant arranged at the vertical and compounds at the horizontal. From this table we obtained the followings: 1) Ligands with vitamin D structure showed similar transactivation pattern each other. 2) Ligands with vitamin D structure have ten essential residues (Y143, D144, L233, 1271, R274, W286, Y295, H397, Y401, F422) for transactivation. 3) Each group of ligands with similar structure showed characteristic activity pattern. 4) Number of essential residues increases for ligands with weak activity. 5) Mutants having stronger activity than the wild type increase in vitamin D super-agonists. 6) LCAs without vitamin D structure show different activity pattern from vitamin D ligands. Based on these results we will investigate discrimination mechanism of vitamin D actions.

我们用二维丙氨酸扫描突变分析(2D-ASMA)方法研究了维生素D受体(VDR)与其配体之间的相互作用。我们对VDR配体结合口袋中所有氨基酸残基的丙氨酸进行了一点突变，并用荧光素酶测定法评价了它们的转录活性。结果显示在二维表上，每个突变体垂直排列，化合物在水平排列。从该表中我们得到如下结果：1)具有维生素D结构的配体之间表现出相似的反式激活模式。2)维生素D结构的配体有10个必需残基(Y143、D144、L233、1271、R274、W286、Y295、H397、Y401、F422)进行反式激活。3)具有相似结构的每一组配体都表现出独特的活性模式。4)活性较弱的配体必需残基数目增加。5)与野生型相比，活性更强的突变体增加了维生素D的超激动剂。6)无维生素D结构的LCA表现出与维生素D配体不同的活性模式。基于这些结果，我们将探讨维生素D作用的区别机制。

项目成果

Sachiko Yamada: "Vitamin D Receptor."(Hochberg Z, editor). Endocr. Dev.: Vitamin D and Rickets KARGER:Basel. 6. 50-68 (2003)

山田幸子：“维生素 D 受体”（Hochberg Z，编辑）。

Mihwa Choi: "Interaction between Vitamin D Receptor and Vitamin D Ligands : Two-Dimensional Alanine Scanning Mutational Analysis."Chem.Biol.. 10. 261-270 (2003)

Mihwa Choi：“维生素 D 受体和维生素 D 配体之间的相互作用：二维丙氨酸扫描突变分析。”Chem.Biol.. 10. 261-270 (2003)

Sachiko Yamada: "Structure-Function Relationships of Vitamin D including Ligand Recognition by the Vitamin D Receptor"Medicinal Research Review. 23. 89-115 (2003)

Sachiko Yamada：“维生素 D 的结构功能关系，包括维生素 D 受体的配体识别”医学研究评论。

Ryutaro Adachi: "Structural Determinants for Vitamin D Receptor Response to Endocrine and Xenobiotic Signals."Mol.Endocrinol. 18. 43-52 (2004)

Ryutaro Adachi：“维生素 D 受体对内分泌和异生信号反应的结构决定因素。”Mol.Endocrinol。

Keiko Yamamoto: "Homology modeling of human 25 -hydroxyvitamin D3 la-hydroxylase (CYP27B 1) based on the crystal structure of rabbit CYP2C5."J. Steroid Biochem. Mol. Biol.. (in press). (2004)

Keiko Yamamoto：“基于兔 CYP2C5 晶体结构的人 25-羟基维生素 D3 1α-羟化酶 (CYP27B 1) 的同源建模。”J.