Covalent labeling of nuclear vitamin D receptor with affinity labeling reagents containing a cross-linking probe at three different positions of the parent ligand: Structural and biochemical implications
Covalent labeling of nuclear vitamin D receptor with affinity labeling reagents containing a cross-linking probe at three different positions of the parent ligand: Structural and biochemical implications
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DOI：
10.1016/j.bioorg.2009.01.001
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发表时间：
2009-02-01
影响因子：
5.1
通讯作者：
Ray, Rahul
中科院分区：
文献类型：
--
作者：
Kaya, Taner;Swamy, Narasimha;Ray, Rahul
维生素D受体（VDR）的结构-功能表征需要鉴定对1 α，25-二羟基维生素D-3（1，25（OH）（2）D-3）的生物学特性重要的VDR-配体结合结构域（VDR-LBD）的结构不同区域。我们假设配体共价连接到VDR-LBD可能会改变“表面结构”的影响配体的生物活性的区域。我们比较了在1，3和11位含有烷基化探针的1，25（OH）（2）D-3的三种亲和烷基化衍生物的抗增殖活性。这些化合物具有与VDR的高亲和性结合;和亲和标记的VDR-LBD。但是，与1，25（OH）（2）D-3相比，只有在3位具有探针的类似物显著改变了角质形成细胞的生长。停靠在VDR-LBD内的这些类似物的分子模型初步鉴定了Ser 237（螺旋-3：1，25（OH）（2）D-3-1-BE）、Cys 288（β-发夹区：1，25（OH）（2）D-3-3-BE）和Tyr 295（螺旋-6：1，25（OH）（2）D-3-11-BE）作为可能被这些试剂修饰的氨基酸。因此，我们得出结论，β-发夹区（由1，25（CH）（2）D-3-3-BE修饰）对1，25（OH）（2）D-3的生长抑制作用最重要，而螺旋3和6对这种活性不太重要。(C)2009 Elsevier Inc. All rights reserved.
Structure-functional characterization of vitamin D receptor (VDR) requires identification of structurally distinct areas of VDR-ligand-binding domain (VDR-LBD) important for biological properties of 1 alpha,25-dihydroxyvitamin D-3 (1,25(OH)(2)D-3). We hypothesized that covalent attachment of the ligand into VDR-LBD might alter 'surface structure' of that area influencing biological activity of the ligand. We compared anti-proliferative activity of three affinity alkylating derivatives of 1,25(OH)(2)D-3 containing an alkylating probe at 1,3 and 11 positions. These compounds possessed high-affinity binding for VDR; and affinity labeled VDR-LBD. But, only the analog with probe at 3-position significantly altered growth in keratinocytes, compared with 1,25(OH)(2)D-3. Molecular models of these analogs, docked inside VDR-LBD tentatively identified Ser237 (helix-3: 1,25(OH)(2)D-3-1-BE), Cys288 (beta-hairpin region: 1,25(OH)(2)D-3-3-BE,) and Tyr295 (helix-6: 1,25(OH)(2)D-3-11-BE,) as amino acids that are potentially modified by these reagents. Therefore, we conclude that the beta-hairpin region (modified by 1,25(CH)(2)D-3-3-BE) is most important for growth inhibition by 1,25(OH)(2)D-3, while helices 3 and 6 are less important for such activity. (C) 2009 Elsevier Inc. All rights reserved.