Studies on Influenza B Virus BM2 Protein Ion Channel

乙型流感病毒BM2蛋白离子通道的研究

基本信息

批准号：
7190559
负责人：
LAWRENCE H PINTO
金额：
$ 27.94万
依托单位：
NORTHWESTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-03-01 至 2010-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7190559
关键词：
Aerosols Affect Amantadine Antiviral Agents Bacteria Bathing Biochemical Biochemistry Biological Cells Cercopithecine Herpesvirus 1 Chemicals Chimeric Proteins Collaborations Coupled Cysteine Cytoplasmic Tail Drug Industry Drug effect disorder Endosomes Epidemic Exposure to Future Genome Human In Vitro Influenza Influenza A virus Influenza A virus M2 protein Influenza B virus Insecta Integral Membrane Protein Iodides Ion Channel Ion Channel Protein Ion Transport Ions Knowledge Laboratories Lipids M2 protein Mass Spectrum Analysis Measures Membrane Molecular Morbidity - disease rate Mutagenesis Mutate Mutation Pathogenicity Peptides Pharmaceutical Preparations Phase Phosphorylation Phosphorylation Site Procedures Property Proteins Protons Reagent Recombinants Relative (related person)Research Personnel Role Scanning Screening procedure Series Serine Simian B disease Structure System Testing Threonine Vesicle Viral Viral Genome Virion Virus Virus Diseases Virus Replication aqueous base design fitness influenzavirus inhibitor/antagonist mortality mutant novel pandemic disease pandemic influenza positional cloning programs protein function reconstitution research study small molecule solid state transmission process

项目摘要

DESCRIPTION (provided by the applicant): Influenza contributes substantially to worldwide morbidity and mortality. Influenza A virus has caused all human pandemics but influenza B virus infections occur annually and in some recent years they have constituted 50% of total influenza virus infections. The relative simplicity of using reverse genetics procedures to alter the genome of influenza A and B viruses to deliberately introduce pathogenicity determinants into the viral genome coupled with the aerosol transmission of influenza viruses, makes the viruses ideal candidates for use by bioterrorists to create epidemics and havoc. Recently, we discovered that the BM2 protein of influenza B virus is an oligomeric integral membrane protein with an ion channel activity. BM2 protein causes acidification of the virion interior that is required for virus uncoating in endosomes. The BM2 protein functions in a manner related to that of the influenza A virus M2 ion channel protein, the latter being the target of the anti-viral drug amantadine: the drug does not inhibit influenza B virus replication or the BM2 ion channel activity. The proposed experiments are designed to elucidate the important features of the structure and function of the BM2 protein and its role in the influenza B virus lifecycle. (1) We will characterize the BM2 protein using biochemistry, mutagenesis and reverse genetics. The phosphorylation state of the BM2 protein will be determined and the effect of specific mutations that affect BM2 function will be determined using reverse genetics procedures to introduce these mutations into influenza B virus. (2) The oligomeric state of the active form of the BM2 protein will be determined using mixtures of BM2 subunits with distinguishable properties. (3) The pore-lining residues of the BM2 channel will be identified using cysteine-scanning mutagenesis followed by biochemical and functional studies. (4) The mechanisms by which the activity of the BM2 protein is modulated will be studied. (5) We will develop systems for measuring the activity of recombinant BM2 protein in vitro in order to provide a useful means for the pharmaceutical industry to screen for antiviral compounds specific for BM2 and to use in biophysical experiments.

描述（由申请人提供）：流感对全世界的发病率和死亡率有很大影响。甲型流感病毒引起了所有人类大流行，但乙型流感病毒感染每年都会发生，近年来，它们已占流感病毒感染总数的 50%。使用反向遗传学程序来改变甲型和乙型流感病毒的基因组，故意将致病性决定因素引入病毒基因组，再加上流感病毒的气溶胶传播，相对简单，使得这些病毒成为生物恐怖分子制造流行病和破坏的理想候选者。。最近，我们发现乙型流感病毒的BM2蛋白是一种具有离子通道活性的寡聚整合膜蛋白。 BM2 蛋白导致病毒粒子内部酸化，这是病毒在内体中脱壳所必需的。 BM2蛋白的功能与甲型流感病毒M2离子通道蛋白的功能相关，后者是抗病毒药物金刚烷胺的靶标：该药物不会抑制乙型流感病毒复制或BM2离子通道活性。所提出的实验旨在阐明 BM2 蛋白结构和功能的重要特征及其在乙型流感病毒生命周期中的作用。 (1) 我们将利用生物化学、诱变和反向遗传学来表征 BM2 蛋白。将确定 BM2 蛋白的磷酸化状态，并使用反向遗传学程序将这些突变引入乙型流感病毒中，确定影响 BM2 功能的特定突变的效果。 (2) BM2 蛋白活性形式的寡聚状态将使用具有可区分特性的 BM2 亚基混合物来确定。 (3) BM2 通道的孔衬残基将通过半胱氨酸扫描诱变进行鉴定，然后进行生化和功能研究。 (4)研究BM2蛋白活性的调节机制。（5）开发重组BM2蛋白体外活性测定系统，为制药工业筛选BM2特异性抗病毒化合物并用于生物物理实验提供有用的手段。