DETERMINATION OF OLIGOMERIC STATE OF HSV GLYCOPROTEINS GD, GH & GL, USING STEM

糖蛋白D(GD)是一种重要的囊膜蛋白。 疱疹。单纯疱疹病毒(HSV)HSV-1和HSV-2都是 含有gD，且gD-1和gD-2在序列上为800/6同源。 这两种蛋白质的截短版本已经在一种 杆状病毒表达系统。这些蛋白质含有所有的氨基 如体外试验所示，功能所必需的酸具有 适当的构象，并且是可溶的。因为它们被截断为 在疏水跨膜区之前的氨基酸306。他们 分别命名为GD-1(306T)和GD-2(306T)。另外两个突变体 Gd-1(306T)基因已被克隆和表达。而野生的则有 三个N-连接的碳水化合物，GD-I(Taat)只有一个，GD I(QAAt) 什么都没有。这两种蛋白质都具有适当的构象和功能 体外试验。Gd-1(306t)蛋白凝胶呈二聚体 过滤数据。STEM中的质量分析与此一致，并且 对所有四个质点的额外测量表明，它们是 都是二聚体。在病毒中发现了单纯疱疹病毒糖蛋白H和L 以杂化齐聚物形式存在的包膜。纯化的Gh/Gl复合体正在进行 在STEM中进行分析以确定其分子量并查看是否 任何结构细节都可以观察到。Gl)(二聚体)/HSV受体 复合体也将使用STEM进行研究。

Glycoprotein D (gD) is an essential envelope protein from the herpes. simplex virus (HSV). The strains HSV-1 and HSV-2 both contain gD, and gD-1 and gD-2 are 800/6 homologous in sequence. Truncated versions of the two proteins have been cloned in a baculovirus expression system. These proteins have all the amino acids necessary for function as shown by in vitro assays, have the proper conformation, and are soluble. since they are truncated at amino acid 306 prior to the hydrophobic transmembrane region. They are called gD-1(306t) and gD-2(306t). Two additional mutants of gD-1(306t) have.been cloned and expressed. Whereas the wild-type has three N-linked carbohydrates, gD-I(TAAt) has only one and gD I (QAAt) has none. Both proteins have the -proper conformation and function in in vitro assays. The protein gD-1(306t) appears dimeric by gel filtration data. Mass analysis in the STEM agrees with this and additional measurements on all four of these protems indicate they are all dimers. The HSV glycoproteins H and L are found in the virus envelope as a hetero-oligomer. The purified gH/gL complex is being analyzed in the STEM to determine its molecular weight and to see if any structural details can be observed. The gl) (dimer)/HSV receptor complex is also going to be studied using the STEM.