Kameyama Y, Yoshioka S, Nozawa Y
Biochim Biophys Acta. 1984 Mar 27;793(1):28-33.
The purpose of the present studies with in vivo isotope labeling was to explore the mechanism by which the fatty acyl composition of membrane phospholipids was altered quickly after a downward temperature-shift in Tetrahymena pyriformis. 1. When 39 degrees C-grown Tetrahymena cells were shifted to 15 degrees C, unsaturated fatty acids, especially gamma-linolenic acid, were increased, with compensating decrease of palmitic acid in phospholipids. 2. However, in the [32P]Pi-prelabeled cells, the specific radioactivities of major membrane phospholipids, phosphatidylethanolamine, phosphatidylcholine and 2-aminoethylphosphonolipid, were not changed within 10 h after shift-down. Furthermore, in the [14C]palmitic acid-prelabeled cells, the specific radioactivities toward phospholipids also did not change after temperature-shift. 3. After the shift, the rate of incorporation of [14C]acetate into fatty acids was reduced to less than one-tenth and the principal fatty acid newly synthesized was linoleic acid, and palmitic and gamma-linolenic acids were also formed to almost same degree. 4. In contrast, when cells prelabeled with both [32P]Pi and [14C]palmitic acid were shifted to 15 degrees C and then linoleic and gamma-linolenic acids, which are known to increase during cold acclimation, were added to the medium, the 14C/32P ratios of major phospholipids were progressively decreased until 5 h after the shift. These results suggest the mechanism for the alteration of phospholipid fatty acyl composition at lowered growth temperature by which preexisting fatty acids of membrane phospholipids would be deacylated and then, after modification to adequate unsaturated fatty acids by desaturation and/or elongation, be reacylated again into lysophospholipids.
本体内同位素标记研究的目的是探索在梨形四膜虫温度下降后膜磷脂的脂肪酰基组成迅速改变的机制。1. 当在39℃生长的梨形四膜虫细胞转移到15℃时,不饱和脂肪酸,尤其是γ-亚麻酸增加,同时磷脂中的棕榈酸相应减少。2. 然而,在[32P]Pi预标记的细胞中,主要膜磷脂磷脂酰乙醇胺、磷脂酰胆碱和2-氨基乙基磷脂的比放射性在温度下降后10小时内没有变化。此外,在[14C]棕榈酸预标记的细胞中,温度变化后磷脂的比放射性也没有变化。3. 温度变化后,[14C]乙酸掺入脂肪酸的速率降至不到十分之一,新合成的主要脂肪酸是亚油酸,棕榈酸和γ-亚麻酸的形成程度也几乎相同。4. 相反,当用[32P]Pi和[14C]棕榈酸预标记的细胞转移到15℃,然后将已知在冷适应过程中增加的亚油酸和γ-亚麻酸添加到培养基中时,主要磷脂的14C/32P比值在转移后5小时内逐渐降低。这些结果提示了在较低生长温度下磷脂脂肪酰基组成改变的机制,即膜磷脂中原有的脂肪酸会被去酰基化,然后在通过去饱和和/或延长作用修饰为适当的不饱和脂肪酸后,再次被重新酰基化到溶血磷脂中。