Kinney A J, Clarkson D T, Loughman B C
Biochem J. 1987 Mar 15;242(3):755-9. doi: 10.1042/bj2420755.
The incorporation of [14C]choline chloride and [14C]glycerol into segments taken from rye (Secale cereale L., cv. Rheidal) roots was greater in segments from roots grown at 5 degrees C than in segments taken from roots growing at 20 degrees C. The incorporation was measured at the temperature at which the root had been growing. Measurements in vitro of the enzymes of the nucleotide pathway showed activity of choline kinase (EC 2.7.1.32), choline-phosphate cytidylyltransferase (EC 2.7.7.15) and cholinephosphotransferase (EC 2.7.8.2) to be higher in homogenates from the cooler roots when assayed at 5 degrees C than the activities assayed at 20 degrees C in the 20 degrees C-root homogenates. Changes in vivo in the pool sizes of the CDP-base intermediates with temperature, relative differences in nucleotide-pathway-enzyme activities and a pulse-chase experiment with [14C]choline indicated that the rate-limiting step for phosphatidylcholine biosynthesis in this tissue, at both temperatures, was the reaction catalysed by cytidylyltransferase.
将[14C]氯化胆碱和[14C]甘油掺入黑麦(Secale cereale L.,品种Rheidal)根部分段中的量,在5摄氏度下生长的根的分段中比在20摄氏度下生长的根的分段中更多。掺入量是在根生长的温度下测定的。对核苷酸途径的酶进行体外测量表明,当在5摄氏度下测定时,来自较冷根的匀浆中胆碱激酶(EC 2.7.1.32)、胆碱磷酸胞苷转移酶(EC 2.7.7.15)和胆碱磷酸转移酶(EC 2.7.8.2)的活性高于在20摄氏度下测定的20摄氏度根匀浆中的活性。CDP - 碱基中间体的库大小随温度的体内变化、核苷酸途径酶活性的相对差异以及用[14C]胆碱进行的脉冲追踪实验表明,在这两个温度下,该组织中磷脂酰胆碱生物合成的限速步骤是由胞苷转移酶催化的反应。