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

EBV 蛋白激酶的结构组织和酶学的定义。

• 批准号: 7636886
• 负责人: Edward Gershburg
• 金额: $ 25.29万
• 依托单位: SOUTHERN ILLINOIS UNIVERSITY SCH OF MED
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-12 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7636886
• 关键词: Address; Affect; Affinity Chromatography; Antiviral Agents; Antiviral Therapy; Attention; Biochemical; Biochemistry; Biological; Biological Assay; Biology; Cell Culture Techniques; Cell Line; Cells; Complement; Complex; DNA biosynthesis; Development; EBV-associated disease; Enzymatic Biochemistry; Enzymes; Epstein-Barr Virus Infections; Family; Figs - dietary; Foundations; Future; Ganciclovir; Genes; Goals; Herpesviridae; Human; Human Herpesvirus 4; Immunoblotting; Immunoprecipitation; In Vitro; Kinetics; Knowledge; Life Cycle Stages; Maps; Measures; Mediating; Molecular Biology; Mutagenesis; Myelin Basic Proteins; Peptides; Pharmacologic Substance; Phosphorylation; Phosphotransferases; Play; Property; Protein Inhibition; Protein Kinase; Proteins; Reaction; Regulation; Role; Screening procedure; Site-Directed Mutagenesis; Structure; Techniques; Testing; Thymidine Kinase; Time; Viral; Viral Proteins; Virus; Virus Diseases; Virus Replication; base; design; effective therapy; in vitro activity; inhibitor/antagonist; interest; mutant; novel; nucleoside analog; overexpression; pathogen; prevent; public health relevance; small molecule; superinfection; viral DNA

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.

描述（由申请人提供）：疱疹病毒编码蛋白激酶（HPK）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，使其成为抗病毒治疗的有吸引力的靶点。该提案的主要目标是研究和开发病毒编码靶标，以开发针对人类病原体 Epstein-Barr 病毒 (EBV) 的新型安全有效的抗病毒药物。该项目采用综合方法：通过分子生物学和生物化学技术研究EBV编码蛋白激酶（EBV-PK）的结构和特性，并探索小分子和基于肽的策略来寻找抑制剂。第一个具体目标是通过诱变研究 EBV-PK 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，本申请旨在表征一种对 EBV 复制至关重要的独特激酶，并鉴定能够有效抑制这种酶的化合物，从而有可能抑制病毒。因此，它解决了一般生物学问题和药学问题。公共卫生相关性鉴于与 EBV 感染相关的疾病数量不断增加，其中许多都非常难以治疗，因此有必要开发和测试 EBV 靶向治疗方法，以尝试设计更好的治疗方法。该提案中概述的研究将增进对 EBV 生物学的了解，并评估开发 EBV 相关疾病有效治疗的新靶点。