Lecithin Biosynthetic Enzymes of Onion Stem and the Distribution of Phosphorylcholine-Cytidyl Transferase among Cell Fractions

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Abstract
Enzymatic activities of the cytidine 5'-diphosphate choline pathway for lecithin biosynthesis were demonstrated in homogenates of onion stem (Allium cepa). Choline kinase activity was present in the postmicrosomal supernatant, with less than 3% sedimenting with the particulate fractions. Phosphorylcholine-cytidyl transferase was distributed among all fractions, and phosphorylcholine-glyceride transferase was predominantly found in the particulate fraction.The phosphorylcholine-cytidyl transferase activity of onion stem required a divalent ion (Mg(2+) or Mn(2+)) for activity, was inhibited by Ca(2+), and was specific for cytidine triphosphate, with optimal activity in the range pH 6 to 7. To evaluate the distribution among cell fractions, conditions of pH, cofactors, substrate, and assay were optimized for each fraction. One-third of the transferase activity sedimented with the mitochondria-proplastids fraction, and one-third was in the microsomal supernatant. The dictyosome fraction contained about 10% of the total activity but showed a greater specific activity than the other fractions. Similar results were obtained with homogenates from rat liver, in that purified Golgi apparatus fractions contained the highest phosphorylcholine-cytidyl transferase activity on a protein basis when compared with other cell fractions at pH 7.2.