Gaillard Stéphanie, Grasfeder Linda L, Haeffele Christiane L, Lobenhofer Edward K, Chu Tzu-Ming, Wolfinger Russ, Kazmin Dmitri, Koves Timothy R, Muoio Deborah M, Chang Ching-Yi, McDonnell Donald P
Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, North Carolina 27710.
Cogenics Division, Clinical Data, Inc., Research Triangle Park, North Carolina 27709.
Mol Cell. 2006 Dec 8;24(5):797-803. doi: 10.1016/j.molcel.2006.10.012.
In the absence of specific high-affinity agonists and antagonists, it has been difficult to define the target genes and biological responses attributable to many of the orphan nuclear receptors (ONRs). Indeed, it appears that many members of this receptor superfamily are not regulated by classical small molecules but rather their activity is controlled by interacting cofactors. Motivated by this finding, we have developed an approach to genetically isolate specific receptor-cofactor pairs in cells, allowing us to define the biological responses attributable to each complex. This is accomplished by using combinatorial peptide phage display to engineer the receptor interacting domain of each cofactor such that it interacts selectively with one nuclear receptor. In this study, we describe the customization of PGC-1alpha and its use to study the biology of the estrogen-related receptor alpha (ERRalpha) in cultured liver cells.
在缺乏特异性高亲和力激动剂和拮抗剂的情况下，很难确定许多孤儿核受体（ONR）所对应的靶基因和生物学反应。实际上，该受体超家族的许多成员似乎不受经典小分子的调控，而是其活性由相互作用的辅因子控制。受这一发现的启发，我们开发了一种在细胞中通过基因手段分离特定受体 - 辅因子对的方法，使我们能够确定每个复合物所对应的生物学反应。这是通过使用组合肽噬菌体展示技术对每个辅因子的受体相互作用结构域进行工程改造来实现的，以便它能选择性地与一种核受体相互作用。在本研究中，我们描述了PGC - 1α的定制及其在培养的肝细胞中用于研究雌激素相关受体α（ERRα）生物学特性的应用。
