Pohjanpelto P, Hölttä E, Jänne O A, Knuutila S, Alitalo K
J Biol Chem. 1985 Jul 15;260(14):8532-7.
We have isolated from an arginase-deficient polyamine-dependent Chinese hamster ovary cell line a new mutant strain that has greatly increased ornithine decarboxylase activity. This enables the cells, in the absence of ornithine, to decarboxylate lysine into cadaverine (diaminopentane) that is further converted into N-(3-aminopropyl)cadaverine and N,N'-bis(3-aminopropyl)cadaverine. These unusual polyamines can support the growth of the cells without added polyamines derived from ornithine. Immunoreactive ornithine decarboxylase-like protein was clearly increased in the mutant cells but could not solely account for the greatly increased enzyme activity. Southern blot analysis of DNA hybridized to a plasmid carrying ornithine decarboxylase-cDNA revealed at least a 32-fold amplification of the ornithine decarboxylase gene. Ornithine decarboxylase-mRNA concentration was also highly increased in the cells. The half-life of the enzyme and the Km for ornithine were not altered from those of the parental cell line.
我们从一种精氨酸酶缺陷型多胺依赖性中国仓鼠卵巢细胞系中分离出一种新的突变株,该突变株的鸟氨酸脱羧酶活性大幅增加。这使得细胞在没有鸟氨酸的情况下,能够将赖氨酸脱羧生成尸胺(二氨基戊烷),尸胺进一步转化为N-(3-氨基丙基)尸胺和N,N'-双(3-氨基丙基)尸胺。这些不同寻常的多胺能够在不添加源自鸟氨酸的多胺的情况下支持细胞生长。突变细胞中免疫反应性鸟氨酸脱羧酶样蛋白明显增加,但这并不能完全解释酶活性的大幅增加。与携带鸟氨酸脱羧酶-cDNA的质粒杂交的DNA的Southern印迹分析显示,鸟氨酸脱羧酶基因至少扩增了32倍。细胞中鸟氨酸脱羧酶-mRNA的浓度也显著增加。该酶的半衰期和对鸟氨酸的Km值与亲代细胞系相比没有改变。