Rose J K, Adams G A, Gallione C J
Proc Natl Acad Sci U S A. 1984 Apr;81(7):2050-4. doi: 10.1073/pnas.81.7.2050.
The transmembrane glycoprotein (G protein) of vesicular stomatitis virus (VSV) is known to contain 1-2 mol of covalently linked fatty acid (palmitate) per mol of protein. G protein is oriented in cellular membranes such that the carboxyl-terminal 29 amino acids protrude into the cytoplasm. We have obtained expression in eukaryotic cells of mutagenized cDNA clones that encode VSV G proteins lacking portions of this cytoplasmic domain. Labeling of these truncated proteins with [3H]palmitate indicated that the palmitate might be linked to an amino acid residue within the first 14 residues on the carboxyl-terminal side of the transmembrane domain. Using oligonucleotide directed mutagenesis, we changed the single codon specifying cysteine in this domain to a codon specifying serine. Expression of this mutant gene results in synthesis of a G protein lacking palmitate. We suggest that linkage of palmitate to G protein is through the cysteine in the cytoplasmic domain and that such a linkage may occur in many viral and cellular glycoproteins. The G protein lacking palmitate is glycosylated and is transported normally to the cell surface.
已知水泡性口炎病毒（VSV）的跨膜糖蛋白（G蛋白）每摩尔蛋白质含有1至2摩尔共价连接的脂肪酸（棕榈酸）。G蛋白在细胞膜中的取向使得羧基末端的29个氨基酸突出到细胞质中。我们已经在真核细胞中获得了诱变的cDNA克隆的表达，这些克隆编码缺少该细胞质结构域部分的VSV G蛋白。用[3H]棕榈酸标记这些截短的蛋白质表明，棕榈酸可能与跨膜结构域羧基末端一侧前14个残基内的一个氨基酸残基相连。使用寡核苷酸定向诱变，我们将该结构域中指定半胱氨酸的单个密码子改变为指定丝氨酸的密码子。该突变基因的表达导致缺乏棕榈酸的G蛋白的合成。我们认为棕榈酸与G蛋白的连接是通过细胞质结构域中的半胱氨酸，并且这种连接可能发生在许多病毒和细胞糖蛋白中。缺乏棕榈酸的G蛋白被糖基化并正常转运到细胞表面。
