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。

基本信息

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
项目类别：
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

项目摘要

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）：由于越来越多的证据表明它们参与了病毒生命周期的不同方面，因此最近引起了疱疹病毒编码的蛋白激酶（HPK）的越来越多的关注，从而使它们对抗病毒药疗法具有吸引力。该提案的广泛目标是研究和利用针对人类病原体爱泼斯坦 - 巴尔病毒（EBV）的新型安全有效抗病毒剂的病毒编码靶标。该项目采用合并的方法：将通过分子生物学和生物化学技术来研究EBV编码的蛋白激酶（EBV-PK）的结构和特性，并将探索小分子和基于肽的策略的抑制剂。第一个具体目的是通过诱变来研究EBV-PK结构，表征其在蛋白质底物和Ganciclovir（GCV）方面的活性，并开发一种用于快速有效筛选抑制剂的测定法。将在体外和细胞培养中研究EBV-PK突变体的活性。第二个特定目的旨在研究小分子和基于肽的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，以抑制病毒复制。因此，该应用旨在表征独特的激酶，该激酶对于EBV复制至关重要，并识别可以有效抑制该酶的化合物，并可能抑制病毒。因此，它解决了一个一般的生物学问题和药物问题。鉴于与EBV感染相关的条件越来越多的公共卫生相关性（众所周知，其中许多条件都难以治疗，对EBV靶向疗法的开发和测试都是有必要的，试图设计更好的治疗方法。该提案中概述的研究将提高对EBV生物学的了解，并评估有效治疗EBV相关疾病的新目标。