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.

我们通过一种新方法研究了维生素D受体（VDR）及其配体之间的相互作用，该方法称为二维丙氨酸扫描突变分析（2D-ASMA）。我们对VDR配体结合袋的所有氨基酸残基的丙氨酸做出了单点突变体，并使用荧光素酶测定方法使用12种配体进行了转录活性。结果呈现在二维表上，每个突变体在垂直方向上排列，并在水平上进行化合物。从该表中我们获得了以下内容：1）具有维生素D结构的配体显示相似的反式激活模式。 2）具有维生素D结构的配体具有十个必需残基（Y143，D144，L233，1271，R274，W286，W286，Y295，H397，Y401，F422）用于反式激活。 3）每组具有相似结构的配体都显示出特征活性模式。 4）活性较弱的配体的基本残基数量增加。 5）与野生型具有更强活性的突变体增加了维生素D超级激动剂。 6）没有维生素D结构的LCA显示与维生素D配体不同的活性模式。基于这些结果，我们将研究维生素D作用的歧视机制。

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)

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。

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。

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.