MacDonald M J, Chang C M
Diabetes. 1985 Mar;34(3):246-50. doi: 10.2337/diab.34.3.246.
Phosphoenolpyruvate carboxykinase activity was measured in rat pancreatic islet cytosol and mitochondria. No carboxykinase activity was detected under a variety of conditions, including those that increase phosphoenolpyruvate carboxykinase activity in nonislet tissues, such as starving animals or incubating the islet extracts with Fe2+ or Mn2+ before assaying for enzyme activity. The amounts of islet cytosol protein used exceeded those of liver in companion assays used as controls. It was calculated that if islet phosphoenolpyruvate carboxykinase activity was 0.005 that of liver, or 1 X 10(-5) as high as pyruvate kinase activity in islets, it should have been detected in the assays used. Ferroactivator is a protein that permits Fe2+ to activate phosphoenolpyruvate carboxykinase and it is ubiquitous to many tissues that do and even do not contain the carboxykinase. Ferroactivator activity was not detectable in pancreatic islets. Pyruvate kinase, an enzyme that catalyzes a reaction that is essentially the opposite of that catalyzed by phosphoenolpyruvate carboxykinase (i.e., phosphoenolpyruvate formation), is plentiful in islet cytosol. Therefore, even if phosphoenolpyruvate carboxykinase activity is present in pancreatic islets, it is so low that it is unlikely that phosphoenolpyruvate formation would be favored and the contribution of the carboxykinase to intracellular carbohydrate metabolism must be quantitatively unimportant.
在大鼠胰岛细胞溶质和线粒体中测定了磷酸烯醇丙酮酸羧激酶的活性。在多种条件下均未检测到羧激酶活性,这些条件包括那些能增加非胰岛组织中磷酸烯醇丙酮酸羧激酶活性的条件,比如饥饿的动物,或者在测定酶活性之前用Fe2+或Mn2+孵育胰岛提取物。在作为对照的同步试验中,所用胰岛细胞溶质蛋白的量超过了肝脏中的量。据计算,如果胰岛磷酸烯醇丙酮酸羧激酶活性是肝脏的0.005倍,或者是胰岛中丙酮酸激酶活性的1×10(-5) 那么高,在所用的测定中应该能够检测到。铁激活剂是一种能使Fe2+激活磷酸烯醇丙酮酸羧激酶的蛋白质,它在许多含有甚至不含有羧激酶的组织中普遍存在。在胰岛中未检测到铁激活剂活性。丙酮酸激酶是一种催化的反应本质上与磷酸烯醇丙酮酸羧激酶催化的反应相反(即生成磷酸烯醇丙酮酸)的酶,在胰岛细胞溶质中含量丰富。因此,即使胰岛中存在磷酸烯醇丙酮酸羧激酶活性,其活性也非常低,以至于生成磷酸烯醇丙酮酸的可能性不大,并且羧激酶对细胞内碳水化合物代谢的贡献在数量上一定不重要。