Kleinau Gunnar, Brehm Mara, Wiedemann Urs, Labudde Dirk, Leser Ulf, Krause Gerd
Leibniz-Institut für Molekulare Pharmakologie, Robert-Rössle-Strasse 10, D-13125 Berlin, Germany.
Mol Endocrinol. 2007 Feb;21(2):574-80. doi: 10.1210/me.2006-0309. Epub 2006 Nov 16.
Comparison between wild-type and mutated glycoprotein hormone receptors (GPHRs), TSH receptor, FSH receptor, and LH-chorionic gonadotropin receptor is established to identify determinants involved in molecular activation mechanism. The basic aims of the current work are 1) the discrimination of receptor phenotypes according to the differences between activity states they represent, 2) the assignment of classified phenotypes to three-dimensional structural positions to reveal 3) functional-structural hot spots and 4) interrelations between determinants that are responsible for corresponding activity states. Because it is hard to survey the vast amount of pathogenic and site-directed mutations at GPHRs and to improve an almost isolated consideration of individual point mutations, we present a system for systematic and diversified sequence-structure-function analysis (http://www.fmp-berlin.de/ssfa). To combine all mutagenesis data into one set, we converted the functional data into unified scaled values. This at least enables their comparison in a rough classification manner. In this study we describe the compiled data set and a wide spectrum of functions for user-driven searches and classification of receptor functionalities such as cell surface expression, maximum of hormone binding capability, and basal as well as hormone-induced Galphas/Galphaq mediated cAMP/inositol phosphate accumulation. Complementary to known databases, our data set and bioinformatics tools allow functional and biochemical specificities to be linked with spatial features to reveal concealed structure-function relationships by a semiquantitative analysis. A comprehensive discrimination of specificities of pathogenic mutations and in vitro mutant phenotypes and their relation to signaling mechanisms of GPHRs demonstrates the utility of sequence-structure-function analysis. Moreover, new interrelations of determinants important for selective G protein-mediated activation of GPHRs are resumed.
建立野生型和突变型糖蛋白激素受体（GPHRs）、促甲状腺激素受体、促卵泡激素受体和促黄体绒毛膜促性腺激素受体之间的比较，以确定参与分子激活机制的决定因素。当前工作的基本目标是：1）根据受体所代表的活性状态差异来区分受体表型；2）将分类后的表型对应到三维结构位置，以揭示3）功能-结构热点，以及4）负责相应活性状态的决定因素之间的相互关系。由于难以全面研究GPHRs上大量的致病突变和定点突变，且难以改进对单个点突变几乎孤立的考量，我们提出了一个用于系统和多样化序列-结构-功能分析的系统（http://www.fmp-berlin.de/ssfa）。为了将所有诱变数据整合到一个集合中，我们将功能数据转换为统一的标度值。这至少能使它们以粗略分类的方式进行比较。在本研究中，我们描述了汇编的数据集以及广泛的功能，用于用户驱动的受体功能搜索和分类，如细胞表面表达、激素结合能力的最大值，以及基础的和激素诱导的Gαs/Gαq介导的环磷酸腺苷/肌醇磷酸积累。与已知数据库互补，我们的数据集和生物信息学工具允许将功能和生化特异性与空间特征联系起来，通过半定量分析揭示隐藏的结构-功能关系。对致病突变和体外突变体表型的特异性及其与GPHRs信号传导机制的关系进行全面区分，证明了序列-结构-功能分析的实用性。此外，恢复了对GPHRs选择性G蛋白介导激活重要的决定因素的新相互关系。
