糖皮质激素受体配体结合域的拮抗和激动形式的三维结构:RU-486诱导一种导致活性拮抗作用的反式构象。
The three-dimensional structures of antagonistic and agonistic forms of the glucocorticoid receptor ligand-binding domain: RU-486 induces a transconformation that leads to active antagonism.
作者信息
Kauppi Björn, Jakob Clarissa, Färnegårdh Mathias, Yang Jie, Ahola Harri, Alarcon Maria, Calles Karin, Engström Owe, Harlan John, Muchmore Steven, Ramqvist Anna-Karin, Thorell Susanne, Ohman Lars, Greer Jonathan, Gustafsson Jan-Ake, Carlstedt-Duke Jan, Carlquist Mats
机构信息
Structure Biology, Karo Bio AB, Novum, SE-141 57 Huddinge, Sweden.
出版信息
J Biol Chem. 2003 Jun 20;278(25):22748-54. doi: 10.1074/jbc.M212711200. Epub 2003 Apr 9.
Here we describe the three-dimensional crystal structures of human glucocorticoid receptor ligand-binding domain (GR-LBD) in complex with the antagonist RU-486 at 2.3 A resolution and with the agonist dexamethasone ligand together with a coactivator peptide at 2.8 A. The RU-486 structure was solved in several different crystal forms, two with helix 12 intact (GR1 and GR3) and one with a protease-digested C terminus (GR2). In GR1, part of helix 12 is in a position that covers the co-activator pocket, whereas in the GR3, domain swapping is seen between the crystallographically identical subunits in the GR dimer. An arm consisting of the end of helix 11 and beyond stretches out from one molecule, and helix 12 binds to the other LBD, partly blocking the coactivator pocket of that molecule. This type of GR-LBD dimer has not been described before but might be an artifact from crystallization. Furthermore, the subunits of the GR3 dimers are covalently connected via a disulfide bond between the Cys-736 residues in the two molecules. All three RU-486 GR-LBD structures show that GR has a very flexible region between the end of helix 11 and the end of helix 12.
在此,我们描述了人糖皮质激素受体配体结合域(GR-LBD)与拮抗剂RU-486形成复合物时分辨率为2.3埃的三维晶体结构,以及与激动剂地塞米松配体和共激活剂肽一起时分辨率为2.8埃的三维晶体结构。RU-486的结构通过几种不同的晶体形式解析得到,其中两种形式的螺旋12完整(GR1和GR3),一种形式的C末端经蛋白酶消化(GR2)。在GR1中,螺旋12的一部分处于覆盖共激活剂口袋的位置,而在GR3中,在GR二聚体中晶体学上相同的亚基之间出现了结构域交换。由螺旋11末端及之后部分组成的一个臂从一个分子伸出,螺旋12与另一个LBD结合,部分阻断了该分子的共激活剂口袋。这种类型的GR-LBD二聚体此前未曾描述过,但可能是结晶过程产生的假象。此外,GR3二聚体的亚基通过两个分子中Cys-736残基之间的二硫键共价连接。所有三种RU- GR-LBD结构均表明,GR在螺旋11末端和螺旋12末端之间有一个非常灵活的区域。