THREE DIMENSIONAL STRUCTURE OF PARAMYXOVIRUS HN

描述(改编自申请人的摘要)：副粘液病毒是 儿童呼吸道疾病的主要原因。长期目标是 开发预防副粘病毒引起的呼吸道疾病的方法。 为了实现这一目标，拟议的研究将侧重于 血凝素-神经氨酸酶(HN)的结构与功能关系 新城疫病毒囊膜糖蛋白的原型 副粘病毒，在病毒附着、神经氨酸酶和促进融合中 活动。这些目标概述了波特纳博士实现目标的战略。 目标1：建议：建立一个高分辨率的三维(3D) HN的结构。目标2：建议：直接确定地点和 受体结合、神经氨酸酶和融合促进的三维结构 HN的活动。 波特纳博士的实验室已经通过悬挂使新城疫病毒HN蛋白结晶 液滴-蒸气扩散法。这些晶体对X射线的衍射程度适中 高分辨率(高达2.6A)，表明它们将适用于 HN三维结构的测定。在目标1中，准备大的 更大的晶体数量将被追求，并有可能增加 并帮助获得和分析重金属衍生品 对解决HN结构至关重要。目标2将决定 直接确定的区域的位置和空间布局 作为受体结合和神经氨酸酶(NA)位置的分析 HN与唾液酸类似物的络合结构。支持性证据 将来自细胞结合和NA活性的突变分析 从包含建议的NA中的定点突变的cDNA中表达 和细胞结合部位，以及受体的分离和序列分析 特异性突变和逃逸突变。突变分析和知识 应提供对位置和位置的评估 与球形头部相关的融合促进结构域的结构 HN地区，从而为药物干预提供了额外的目标。

DESCRIPTION (adapted from applicant's abstract): Paramyxoviruses are the major cause of respiratory disease in children. The long-term objective is to develop means to prevent respiratory disease caused by paramyxoviruses. To reach that objective, the proposed research will focus on structure-function relationships of the hemagglutinin-neuraminidase (HN) envelope glycoprotein of Newcastle disease virus (NDV), a prototypic paramyxovirus, in virus attachment, neuraminidase and fusion-promoting activity. The aims outline Dr. Portner's strategy to achieve his goals. Aim 1: Proposal: To establish a high resolution three-dimensional (3-D) structure of HN. Aim 2: Proposal: To determine directly the location and 3-D structure of the receptor binding, neuraminidase, and fusion promoting activities of HN. Dr. Portner's laboratory has crystallized the NDV HN protein by the hanging drop-vapor diffusion method. These crystals diffract X-rays at moderately high resolution (up to 2.6A) indicative that they will be suitable for determination of the HN 3-D structure. In Aim 1, the preparation of large amounts of larger crystals will be pursued with the prospect of increasing the resolution and to aid in obtaining and analyzing heavy metal derivatives essential to the solution of the HN structure. Aim 2 will determine directly the location and spacial arrangement of regions tentatively defined as the receptor binding and neuraminidase (NA) sites by analyzing the 3-D structure of HN complexed with sialic acid analogues. Supporting evidence will come from mutational analysis of cell binding and NA activities expressed from cDNAs that contain site-directed mutations in the proposed NA and cell binding sites, and isolation and sequence analysis of receptor specificity mutations and escape mutants. Mutational analysis and knowledge of the 3-D structure should provide an assessment of the location and structure of the fusion promoting domain associated with the globular head region of HN, thus providing additional targets for drug intervention.