Definition of structural organization and enzymology of the EBV protein kinase.

描述（由申请人提供）：疱疹病毒编码蛋白激酶（HPKs）最近引起了越来越多的关注，因为越来越多的证据表明它们参与病毒生命周期的不同方面，使它们成为抗病毒治疗的有吸引力的靶点。本研究的主要目标是研究和开发一种新型的安全有效的抗病毒药物，以对抗人类病原体eb病毒（EBV）。该项目采用综合方法：ebv编码蛋白激酶（EBV-PK）的结构和性质将通过分子生物学和生物化学技术进行研究，并在寻找抑制剂时，将探索基于小分子和肽的策略。第一个具体目标是通过诱变研究EBV-PK的结构，表征其在蛋白质底物和更昔洛韦（GCV）方面的活性，并开发一种快速有效筛选抑制剂的方法。EBV-PK突变体的活性将在体外和细胞培养中进行研究。第二个具体目标是研究小分子和肽类抑制剂对激酶的抑制作用。选定的EBV-PK抑制剂将在细胞培养中进一步测试其抑制病毒复制的能力。因此，本应用程序旨在表征一种独特的激酶，这对EBV复制至关重要，并确定能够有效抑制这种酶的化合物，从而潜在地抑制病毒。因此，它解决了一般的生物学问题和药学问题。鉴于与eb病毒感染相关的疾病越来越多，其中许多是众所周知的难以治疗，开发和测试eb病毒靶向治疗方法是有必要的，以尝试设计更好的治疗方法。本提案中概述的研究将促进对eb病毒生物学的理解，并为开发有效治疗eb病毒相关疾病的新靶点进行评估。

DESCRIPTION (provided by applicant): Herpesvirus-encoded protein kinases (HPKs) have attracted increasing attention lately due to the growing evidence of their involvement in different aspects of the viral life cycle, making them appealing targets for antiviral therapy. The broad objectives of this proposal are to study and exploit a virally encoded target for novel safe and effective antiviral agents against the human pathogen Epstein-Barr virus (EBV). The project takes a combined approach: the structure and properties of the EBV-encoded protein kinase (EBV-PK) will be studied by molecular biology and biochemistry techniques, and in search for the inhibitors both small-molecule and peptide-based strategies will be explored. The first specific aim is to study EBV-PK structure by mutagenesis, characterize its activity in regard to protein substrates and ganciclovir (GCV) and develop an assay for fast and effective screening of inhibitors. The activity of EBV-PK mutants will be studied both in vitro and in cell culture. The second specific aim is designed to study inhibition of the kinase by small-molecule and peptide-based inhibitors. Selected EBV-PK inhibitors will be further tested in cell culture for their ability to inhibit viral replication. Thus, this application aims to characterize a unique kinase, which is crucial for EBV replication, and to identify compounds that will efficiently inhibit this enzyme, potentially inhibiting the virus. Therefore it addresses a general biological question and a pharmaceutical problem. PUBLIC HEALTH RELEVANCE Given the growing number of conditions associated with EBV infection, many of which are notoriously difficult to treat, the development and testing of EBV-targeting therapy approaches is warranted in attempts to devise better treatments. The studies outlined in this proposal will advance understanding of EBV biology and evaluate a novel target for the development of the effective treatment of EBV-associated diseases.