Baquet A, Gaussin V, Bollen M, Stalmans W, Hue L
Hormone and Metabolic Research Unit, International Institute of Cellular and Molecular Pathology, Brussels, Belgium.
Eur J Biochem. 1993 Nov 1;217(3):1083-9. doi: 10.1111/j.1432-1033.1993.tb18340.x.
The activation of hepatic glycogen synthase by the amino-acid-induced cell swelling has been attributed to the stimulation of [glycogen-synthase]-phosphatase resulting from an increase in the intracellular content in glutamate and aspartate, and a decrease in intracellular Cl-, which is a compensatory response to cell swelling [Meijer, A. J., Baquet, A., Gustafson, L., van Woerkom, G. M. & Hue, L. (1992) J. Biol. Chem. 267, 5823-5828]. Here we studied whether the activation of acetyl-CoA carboxylase by cell swelling could be explained by the same mechanism. The activation of endogenous or purified acetyl-CoA carboxylase was measured in gel-filtered liver extracts or cytosols. No activation could be observed under basal conditions but a fivefold stimulation was obtained with concentrations of glutamate (20-25 mM) found in hepatocytes incubated with glutamine. A similar stimulation was also observed with other dicarboxylic acids such as malonate and succinate, or with metal ions like Mg2+, Ca2+ and Mn2+ (10 mM). The addition of 50-100 mM Cl- was found to inhibit the activation of acetyl-CoA carboxylase by some 20-30%. Mg2+ was also found to stimulate the activation of the endogenous glycogen synthase. The glutamate-stimulated and Mg(2+)-stimulated activation of glycogen synthase and acetyl-CoA carboxylase was unaffected by 10 microM inhibitor-2, a specific inhibitory protein of protein phosphatase-1, but could be nearly completely blocked by the phosphatase inhibitor microcystin-LR. Our data suggest that the amino-acid-induced activation of acetyl-CoA carboxylase and glycogen synthase in the liver occurs by a common ionic mechanism.
氨基酸诱导的细胞肿胀对肝糖原合酶的激活作用,被认为是由于谷氨酸和天冬氨酸细胞内含量增加以及细胞内氯离子减少,从而刺激了[糖原合酶]-磷酸酶,这是对细胞肿胀的一种代偿性反应[梅杰,A. J.,巴凯,A.,古斯塔夫森,L.,范·沃科姆,G. M. & 于埃,L.(1992年)《生物化学杂志》267,5823 - 5828]。在此,我们研究了细胞肿胀对乙酰辅酶A羧化酶的激活作用是否可用相同机制来解释。在内源或纯化的乙酰辅酶A羧化酶的激活实验中,使用凝胶过滤的肝提取物或胞质溶胶进行测量。在基础条件下未观察到激活现象,但在与谷氨酰胺一起孵育的肝细胞中发现的谷氨酸浓度(20 - 25 mM)下,激活作用增强了五倍。在用其他二羧酸如丙二酸和琥珀酸,或金属离子如Mg2 +、Ca2 +和Mn2 +(10 mM)处理时,也观察到了类似的激活作用增强。发现添加50 - 100 mM Cl - 可使乙酰辅酶A羧化酶的激活作用受到约20 - 30%的抑制。还发现Mg2 +可刺激内源性糖原合酶的激活。糖原合酶和乙酰辅酶A羧化酶的谷氨酸刺激的激活作用以及Mg(2 +)刺激的激活作用不受10 μM抑制剂 - 2(蛋白磷酸酶 - 1的一种特异性抑制蛋白)的影响,但可被磷酸酶抑制剂微囊藻毒素 - LR几乎完全阻断。我们的数据表明,肝脏中氨基酸诱导的乙酰辅酶A羧化酶和糖原合酶的激活是通过一种共同的离子机制发生的。