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

糖蛋白D（Glycoprotein D，gD）是一种重要的包膜蛋白， 疱疹 单纯病毒（HSV）。 HSV-1和HSV-2两种病毒株 gD-1和gD-2在序列上有800/6的同源性。 这两种蛋白质的截短版本已经在一个 杆状病毒表达系统 这些蛋白质含有所有的氨基 如体外测定所示，功能所必需的酸具有 正确的构象，并且是可溶的。 因为它们被截断， 在疏水跨膜区之前的氨基酸306。 他们 被称为gD-1（306 t）和gD-2（306 t）。 另外两个突变体 gD-1（306 t）已被克隆和表达。 而野生型 三个N-连接的碳水化合物，gD-I（TAAt）只有一个，gD I（QAAt） 没有。 这两种蛋白质都具有适当的构象和功能， 体外测定。 凝胶电泳显示gD-1（306 t）蛋白为二聚体 过滤数据 STEM中的质量分析同意这一点， 对所有四个保护的额外测量表明它们是 都是二聚体。 HSV糖蛋白H和L存在于病毒中， 包封为杂低聚物。 纯化的gH/gL复合物被 在STEM中进行分析，以确定其分子量， 可以观察到任何结构细节。 gl）（二聚体）/HSV受体 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.