Wen D, Ding M X, Schlesinger M J
Virology. 1986 Aug;153(1):150-4. doi: 10.1016/0042-6822(86)90016-4.
Saccharomyces cerevisiae strains transformed with plasmids containing cDNAs coding for the glycoproteins of vesicular stomatitis or Sindbis viruses can be induced to produce large amounts of glycosylated virus glycoproteins. Studies reported here show that these proteins from high molecular weight disulfide-linked oligomers in the yeast endoplasmic reticulum. Oligomers were also found for two genetically altered forms of VSV G; one of these was lacking the membrane anchor domain and the other had the cysteine in the cytoplasmic tail replaced with serine. These oligomers can be separated from the bulk of yeast proteins by brief high-speed centrifugation of yeast extracts prepared by boiling cells with 1% sodium dodecyl sulfate. Treatment with thiol-reducing agents converts the oligomers to soluble monomeric forms, and this procedure leads to a substantial purification of glycoproteins from bulk yeast protein.
用含有编码水泡性口炎病毒或辛德毕斯病毒糖蛋白的cDNA的质粒转化的酿酒酵母菌株，可被诱导产生大量糖基化的病毒糖蛋白。此处报道的研究表明，这些蛋白质在酵母内质网中形成高分子量的二硫键连接的寡聚体。还发现了水泡性口炎病毒糖蛋白（VSV G）的两种基因改变形式的寡聚体；其中一种缺少膜锚定结构域，另一种细胞质尾巴中的半胱氨酸被丝氨酸取代。通过用1%十二烷基硫酸钠煮沸细胞制备的酵母提取物进行短暂高速离心，可以将这些寡聚体与大部分酵母蛋白分离。用硫醇还原剂处理可将寡聚体转化为可溶性单体形式，此过程可从大量酵母蛋白中大量纯化糖蛋白。
