Molecular basis of protein-ligand interactions.

这项研究计划的长期目标是了解蛋白质-配体相互作用的分子机制和功能后果。我们对与金属离子和/或其他蛋白质相互作用的蛋白质特别感兴趣，因为我们对蛋白质如何识别这些配体的理解仍然存在重要的空白。病毒蛋白特别适合于这些研究，因为单个蛋白经常含有许多不同的酶活性，因此可以与各种配体相互作用。在目前的研究项目中，我们的重点是来自丙型肝炎病毒（HCV）的NS5A蛋白。我们相信了解NS5A的功能可以提高对HCV感染和病毒传播的认识。*** ***在细胞中发现了各种各样的金属离子。对于大多数蛋白质来说，金属离子的存在对其活性/功能至关重要。在NS5A蛋白的晶体结构中发现了锌原子，锌原子似乎对该蛋白的功能和病毒复制都至关重要。然而，锌原子在蛋白质中的确切作用仍然是未知的，其功能作用尚未得到令人满意的解释，仍然是一个争论和猜测的问题。虽然目前还没有发现NS5A的酶促功能，但它是HCV复制酶复合体的重要组成部分，对细胞通路和过程有广泛的影响，包括先天免疫、宿主细胞生长和增殖。NS5A最令人兴奋的研究领域之一是它与宿主细胞蛋白的相互作用及其调节宿主途径的能力。幸运的是，各种细胞结合伙伴已经被确定。我们相信，更好地了解NS5A与细胞蛋白之间的相互作用将揭示病毒诱导的细胞通路调节的关键特征。******我们这项研究计划的长期目标是了解决定酶及其配体之间相互作用的分子决定因素，并了解其功能后果。为了实现这些目标，我们目前的具体目标是：******1。表征金属离子结合HCV NS5A蛋白的热力学和结构后果；***2。2 .研究细胞蛋白与NS5A蛋白的相互作用及其功能影响。探讨NS5A对细胞基因表达的调控能力。******研究生和本科生都将是我们研究项目的驱动力。通过在分子生物学、生物化学和结构生物学领域的训练，他们将继续为该领域做出原创性的贡献。显然，我们的首要任务是利用国家科学研究委员会的资助，为有兴趣从事有回报的研究事业的学生提供优秀的培训。

The long-term objective of this research program is to understand the molecular mechanisms and functional consequences of proteins-ligand interactions. We are especially interested in proteins that interact with metal ions and/or other proteins because there are still important gaps in our understanding of how proteins recognize these ligands. Viral proteins are especially well suited for these studies because a single protein frequently harbors many different enzymatic activities and can therefore interact with various ligands. In the current research program, we are focusing on the NS5A protein from the Hepatitis C virus (HCV). We believe that understanding the functions of NS5A could lead to an improved knowledge of HCV infection and viral propagation.*** ***A wide variety of metal ions are found in cells. For most proteins, the presence of metal ions is crucial for activity/function. A zinc atom is found in the crystal structure of the NS5A protein and appears critical for both the function of the protein and for viral replication. However, the precise role of this zinc atom in the protein is still unknown and its functional role has not yet received a satisfactory explanation and is still a matter of debate and speculation. While no known enzymatic function has been ascribed to NS5A, it is an essential component of the HCV replicase complex and exerts a wide range of effects on cellular pathways and processes, including innate immunity, host cell growth and proliferation. One of the most exciting areas of research on NS5A is its interactions with host cell proteins and its ability to modulate host pathways. Fortunately, various cellular binding partners have already been identified. We believe that a better understanding of the interactions between NS5A and cellular proteins will reveal key features in the virus-induced modulation of cellular pathways.******Our long-term objective for this research program is to understand the molecular determinants that dictate the interactions between enzymes and their ligands, and to understand their functional consequences. To achieve these goals, our current specific objectives are:******1. To characterize the thermodynamic and structural consequences of metal ion binding to the HCV NS5A protein;***2. To characterize the interaction between cellular proteins and the NS5A protein, and their functional consequences.***3. To investigate the ability of NS5A to modulate the expression of cellular genes.******Both graduate and undergraduate students will be the driving force in our research program. Through their training in the fields of molecular biology, biochemistry, and structural biology they will continue to make original contributions to the field. Clearly, it is a priority for us to use the NSERC grant to provide excellent training for students interested in rewarding careers in research.