Polokoff M A, Wing D C, Raetz C R
J Biol Chem. 1981 Aug 10;256(15):7687-90.
An in situ autoradiographic assay for CDP-ethanolamine:1,2-sn-diacylglycerol ethanolamine phosphotransferase (EC 2.7.8.1) activity in Chinese hamster ovary cells was developed and used to screen approximately 10,000 individual mutagen-treated colonies attached to filter paper (Esko, J. D., and Raetz, C. R. H. (1978) Proc. Natl. Acad. Sci. U. S. A. 75, 1190-1193). A variant (strain 40.11) was isolated in which the ethanolamine phosphotransferase specific activity in vitro was 6-10-fold less than in the parent, but the level of CDP-choline:1,2-sn-diacylglycerol choline phosphotransferase (EC 2.7.8.2) activity was normal. In extracts, the mutant was also defective in the synthesis of ethanolamine plasmalogen. In vivo, the short term kinetics of labeling with [32P]phosphate or [14C]ethanolamine was correspondingly altered. However, the long tem growth rate and steady state phospholipid compositions of the mutant and parent were quite similar. These results show that the ethanolamine and choline phosphotransferases of Chinese hamster ovary cells are distinct as judged by genetic criteria, while the biosynthesis of phosphatidylethanolamine and its plasmalogen share common enzymatic component(s).
我们建立了一种用于检测中国仓鼠卵巢细胞中CDP - 乙醇胺：1,2 - 二酰基甘油乙醇胺磷酸转移酶（EC 2.7.8.1）活性的原位放射自显影测定法，并用于筛选附着在滤纸上的约10,000个经诱变处理的单个菌落（埃斯科，J. D.，和雷茨，C. R. H.（1978年）美国国家科学院院刊75，1190 - 1193）。分离出一个变体（40.11株），其体外乙醇胺磷酸转移酶的比活性比亲本低6 - 10倍，但CDP - 胆碱：1,2 - 二酰基甘油胆碱磷酸转移酶（EC 2.7.8.2）的活性水平正常。在提取物中，该突变体在乙醇胺缩醛磷脂的合成方面也存在缺陷。在体内，用[32P]磷酸盐或[14C]乙醇胺标记的短期动力学相应地发生了改变。然而，该突变体和亲本的长期生长速率和稳态磷脂组成非常相似。这些结果表明，从遗传学标准判断，中国仓鼠卵巢细胞的乙醇胺和胆碱磷酸转移酶是不同的，而磷脂酰乙醇胺及其缩醛磷脂的生物合成共享共同的酶成分。
