Methyl TROSY of alanine residues in large protein complexes: development and application

DNA是生命的基石，包含细胞制造蛋白质所需的所有信息。RNA序列中存在一种中间语言，它将基因的信息翻译成蛋白质的氨基酸序列。产生RNA的过程称为转录，是从简单细菌到人类的生物体中基因表达的最重要调控步骤。蛋白质RNA聚合酶（RNAP）是催化转录的酶，并且是大多数基因表达调节的直接或间接靶点。构成我们身体的细胞是高度调节和动态的系统。在细胞内部和周围，功能由不同类型的生物分子（如蛋白质）的相互作用决定。通过在原子水平上识别特定的互补表面，这些相互作用是可能的。为了更好地理解这些细胞机制，研究人员需要开发新技术来研究生物分子。特别是，现代分子技术如核磁共振（NMR）提供了一种强大的方法来确定分子的不同形状（或构象）和运动（或动力学）以及它们之间的相互作用。在这项提案中，我们的目标是开发一种新的NMR应用，可以应用和分析非常大的生物分子，如RNAP。这些技术提供的结构细节也足以阐明酶机制（例如，合成和降解新分子所需的一系列步骤），并分析生物分子的相互作用。我们将利用这种新的方法来了解更多关于病毒如何干扰RNAP的活性，以改变基因表达来满足自己的需要

DNA is the building block of the life and contains all the information that the cells require to manufacture proteins. An intermediate language exists in the sequence of RNA that translates a gene's message into a protein's amino acid sequence. The process of producing RNA is called transcription and is the most important regulatory step in gene expression in organisms ranging from simple bacteria to humans. The protein RNA polymerase (RNAP) is the enzyme which catalyses transcription and is the target, directly or indirectly, of most regulation of gene expression. The cells that make up our bodies are highly regulated and dynamic systems. In and around cells, functions are determined by the interplay of different types of biomolecules, such as proteins. These interactions are possible through the recognition of specific, complementary surfaces at the atomic level. For a better understanding of these cellular mechanisms, researchers need to develop new technologies for the study of biomolecules. In particular, modern molecular techniques like nuclear magnetic resonance (NMR) offer a powerful method to determine the different shapes (or conformations) and motion (or dynamics) of molecules and the interactions between them. In this proposal we aim to develop a novel application of NMR that can be applied and analysed on very large biomolecules, such as RNAP. The structural detail provided by these techniques is also sufficient for the elucidation of enzymatic mechanisms (e.g. the series of steps required for the synthesis and degradation of new molecules) and to analyse the interaction of biomolecules. We will exploit this new methodology to understand more about how viruses can interfere with the activity for the RNAP in order to alter gene expression to serve its own needs