THREE DIMENSIONAL STRUCTURE OF PARAMYXOVIRUS HN

描述（改编自申请人的摘要）：副粘病毒是 儿童呼吸道疾病的主要原因。 长期目标是 研究预防副粘病毒引起的呼吸道疾病的方法。 为达致这个目标，拟议的研究将集中于 血凝素-神经氨酸酶（HN）的结构-功能关系 纽卡斯尔病毒（NDV）的囊膜糖蛋白， 副粘病毒，在病毒附着，神经氨酸酶和融合促进 活动 这些目标概述了波特纳博士实现目标的策略。 目标1：建议：建立一个高分辨率的三维（3-D） HN的结构。 目标2：建议：直接确定地点， 受体结合、神经氨酸酶和融合促进的三维结构 的活动。 博士Portner的实验室已经通过悬挂使NDV HN蛋白结晶， 滴汽扩散法 这些晶体在中等强度下吸收X射线， 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，制备大的 将追求更大的晶体量，其前景是增加 分离并帮助获得和分析重金属衍生物 这对解决HN结构至关重要。 目标2将决定 直接影响到暂定区域的位置和空间布局 作为受体结合和神经氨酸酶（NA）的网站，通过分析的3-D HN与唾液酸类似物复合的结构。 支持证据 将来自细胞结合和NA活性的突变分析 从在提出的NA中含有定点突变的cDNA表达 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 三维结构的评估应提供位置和 与球状头相关的融合促进结构域的结构 HN区域，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.