Selective inhibition of isoprenylation of 21-26-kDa proteins by the anticarcinogen d-limonene and its metabolites.
Selective inhibition of isoprenylation of 21-26-kDa proteins by the anticarcinogen d-limonene and its metabolites.
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发表时间：
1991-09
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通讯作者：
Pamela L. Crowell;R. Chang;Zhibin Ren;Charles E. Elson;Michael N. Gould
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作者：
Pamela L. Crowell;R. Chang;Zhibin Ren;Charles E. Elson;Michael N. Gould
柠檬烯对化学诱导的大鼠乳腺癌具有化疗活性，其中许多乳腺癌含有激活的 ras 基因。鉴于最近发现甲羟戊酸途径中的中间体对 p21ras 和其他细胞生长相关蛋白进行翻译后修饰，以及柠檬烯和这些异戊二烯产物的共同生化起源，我们研究了柠檬烯对蛋白质异戊二烯化的影响。将NIH3T3和人乳腺上皮细胞与洛伐他汀和[2-14C]甲瓦龙酸内酯在不存在和存在柠檬烯的情况下一起孵育。然后对标记的蛋白质进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和荧光照相。在两种细胞系中，柠檬烯以剂量依赖性方式抑制一类 21-26 kDa 细胞蛋白（包括 p21ras 和可能的其他小 GTP 结合蛋白）的异戊二烯化。相比之下，柠檬烯不影响其他几种蛋白质的异戊二烯化，包括核纤层蛋白。柠檬烯在体内广泛代谢，但在培养细胞中不代谢。柠檬烯的两种主要大鼠血清代谢物紫苏酸和二氢紫苏酸在抑制异戊二烯化方面比柠檬烯更有效。这些结果表明，柠檬烯选择性抑制甲羟戊酸途径中 3-羟基-3-甲基戊二酰辅酶 A 还原酶远端的 21-26-kDa 蛋白质的异戊二烯化，并为其化疗活性提供了合理的解释。抑制 p21ras 和其他小 GTP 结合蛋白等蛋白质的异戊二烯化会改变其细胞内定位，从而破坏其生物活性。
Limonene has chemotherapeutic activity against chemically induced rat mammary carcinomas, many of which contain activated ras genes. Given the recent discovery of the post-translational modification of p21ras and other cell growth-associated proteins by intermediates in the mevalonic acid pathway, and the common biochemical origins of limonene and these isoprene products, we investigated the effect of limonene on protein isoprenylation. NIH3T3 and human mammary epithelial cells were incubated with lovastatin and [2-14C]mevalonolactone in the absence and presence of limonene. Labeled proteins were then subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. Limonene inhibited isoprenylation of a class of cellular proteins of 21-26 kDa, including p21ras and possibly other small GTP-binding proteins, in a dose-dependent manner in both cell lines. In contrast, limonene did not affect the isoprenylation of several other proteins, including nuclear lamins. Limonene is metabolized extensively in vivo but not in cultured cells. The two major rat serum metabolites of limonene, perillic acid and dihydroperillic acid, were more potent than limonene in the inhibition of isoprenylation. These results demonstrate that limonene selectively inhibits isoprenylation of 21-26-kDa proteins at a point in the mevalonic acid pathway distal to 3-hydroxy-3-methylglutaryl coenzyme A reductase, and they provide a plausible explanation for its chemotherapeutic activity. Inhibition of isoprenylation of proteins such as p21ras and other small GTP-binding proteins would alter their intracellular localization and, hence, disrupt their biological activity.