Petronini P G, Tramacere M, Wheeler K P, Borghetti A F
Istituto di Patologia Generale, Università degli Studi di Parma, Italy.
Biochim Biophys Acta. 1990 Jul 12;1053(2-3):144-50. doi: 10.1016/0167-4889(90)90006-y.
The activity of amino acid transport System A in avian fibroblasts was increased following incubation of the cells in a medium in which most of the NaCl normally present had been isoosmotically replaced by sucrose. This increase was detectable after 2 h of incubation, reached a maximum at about 4 h, and remained constant thereafter. Transfer of treated cells back to a normal medium resulted in decay of the induced transport activity, with a half-life of less than 2 h. Kinetic analysis revealed that the increase in transport activity arose from an increase in Vmax, with little change in Km. This induction of System A activity did not occur if an inhibitor of either RNA or protein synthesis was present in the modified medium. The use of various different solutes as replacements for NaCl in the incubation medium showed that, although each replacement caused a decrease in both cellular Na+ content and protein synthesis, only disaccharides produced the increase in amino acid transport activity. In addition, estimates of cell volume indicated that, even under iso-osmotic conditions, incubation in the sucrose-containing medium caused initial cell shrinkage, followed by swelling. It is concluded that this induction of System A activity is associated with a volume regulatory process and that this process probably accounts for the parallel responses previously observed when cells were incubated in hyperosmolar media. Induction of amino acid transport activity by this process is distinct from adaptive regulation, caused by amino acid starvation; but the two processes are not strictly additive, and so appear to converge at some step.
将禽成纤维细胞置于一种培养基中培养,其中大部分正常存在的氯化钠已被等渗蔗糖替代,之后细胞中氨基酸转运系统A的活性增加。孵育2小时后可检测到这种增加,在约4小时时达到最大值,此后保持恒定。将处理过的细胞转移回正常培养基会导致诱导的转运活性衰减,半衰期小于2小时。动力学分析表明,转运活性的增加源于Vmax的增加,而Km变化不大。如果在改良培养基中存在RNA或蛋白质合成抑制剂,则不会发生系统A活性的这种诱导。在孵育培养基中使用各种不同的溶质替代氯化钠表明,虽然每种替代物都会导致细胞内钠离子含量和蛋白质合成减少,但只有二糖会使氨基酸转运活性增加。此外,细胞体积估计表明,即使在等渗条件下,在含蔗糖的培养基中孵育也会导致细胞先收缩,随后肿胀。得出的结论是,系统A活性的这种诱导与体积调节过程有关,并且该过程可能解释了之前在高渗培养基中孵育细胞时观察到的平行反应。通过该过程诱导氨基酸转运活性与由氨基酸饥饿引起的适应性调节不同;但这两个过程并非严格相加,因此似乎在某个步骤会趋同。