Differential prenylation of proteins as a function of mevalonate concentration in CHO cells.
Differential prenylation of proteins as a function of mevalonate concentration in CHO cells.
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DOI：
10.1006/abbi.1993.1135
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发表时间：
1993-03
影响因子：
3.9
通讯作者：
H. Rilling;E. Bruenger;L. M. Leining;J. Buss;W. W. Epstein-W.
中科院分区：
文献类型：
--
作者：
H. Rilling;E. Bruenger;L. M. Leining;J. Buss;W. W. Epstein-W.
[5-3H]甲羟戊酸掺入异戊二烯化蛋白质和聚异戊二烯脂质中已被确定为甲羟戊酸合成受到抑制的中国仓鼠卵巢（CHO）细胞中甲羟戊酸浓度的函数。甲羟戊酸进入类异戊二烯代谢的不同终产物的相对掺入明显取决于培养基中甲羟戊酸的浓度。在甲羟戊酸浓度较高时，胆固醇的合成占主导地位，而在较低浓度下，则有利于较高分子量的类异戊二烯的合成。甲羟戊酸与异戊二烯化蛋白质的不同异戊二烯半胱氨酸的相对掺入取决于甲羟戊酸浓度，其中香叶基香叶基半胱氨酸在较高浓度下是主要产物。在低水平的甲羟戊酸法尼基半胱氨酸合成中占主导地位并且检测到香叶基半胱氨酸。来自已用不同浓度的[3H]甲羟戊酸放射性标记的CHO细胞的蛋白质的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳在荧光照相上具有不同的图案，在低浓度下标记的蛋白质相对较少。对特定蛋白质 Ras 的异戊二烯半胱氨酸的影响的研究表明，与大量蛋白质相比，对甲羟戊酸浓度的敏感性要低得多。这些结果表明异戊二烯化的特定蛋白质取决于二磷酸异戊二烯基底物的可用性。
The incorporation of [5-3H]mevalonate into prenylated proteins and polyisoprenoid lipids has been determined as a function of mevalonate concentration in Chinese hamster ovary (CHO) cells that are inhibited in mevalonate synthesis. The relative incorporation of mevalonate into the different end products of isoprenoid metabolism was markedly dependent upon the concentration of mevalonate in the medium. The synthesis of cholesterol was dominant at higher concentrations of mevalonate while higher molecular weight isoprenoids were favored at the lower concentrations. The relative incorporation of mevalonate into the different prenylcysteines of prenylated proteins was dependent upon mevalonate concentration with geranylgeranylcysteine being the principal product at higher concentrations. At low levels of mevalonate farnesylcysteine synthesis predominated and geranylcysteine was detected. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins from CHO cells that had been radiolabeled at different concentrations of [3H]mevalonate had different patterns on fluorography with relatively few proteins labeled at low concentrations. A study of this effect on the prenylcysteines of a specific protein, Ras, showed considerably less sensitivity to mevalonate concentration than bulk protein. These results indicate that the specific proteins that are prenylated depend upon the availability of the isoprenyl diphosphate substrates.