Murphy G, Walker D G
Biochem J. 1974 Oct;144(1):149-60. doi: 10.1042/bj1440149.
A homogeneous preparation of ;malic' enzyme (EC 1.1.1.40) from livers of thyroxine-treated rats was used to prepare in rabbits an antiserum to the enzyme that reacts monospecifically with the ;malic' enzyme in livers of rats in several physiological states. Changes in enzyme activity resulting from modification of the state of the animal are hence due to an altered amount of enzyme protein. The antiserum has been used to precipitate out ;malic' enzyme from heat-treated supernatant preparations of livers from both adult and neonatal rats, in a number of physiological conditions, that had been injected 30min earlier with l-[4,5-(3)H]leucine. The low incorporations of radioactivity into the immunoprecipitable enzyme have permitted the qualitative conclusion that changed enzyme activity in adult rats arises mainly from alterations in the rate of enzyme synthesis. The marked increase in ;malic' enzyme activity that occurs naturally or as a result of thyroxine treatment of the weanling rat is likewise due to a marked increase in the rate of enzyme synthesis possibly associated with a concurrent diminished rate of enzyme degradation.
用甲状腺素处理过的大鼠肝脏中制备出的均一的“苹果酸”酶(EC 1.1.1.40)制剂,在兔子体内制备了针对该酶的抗血清,该抗血清能与处于几种生理状态的大鼠肝脏中的“苹果酸”酶发生单特异性反应。因此,动物状态改变引起的酶活性变化是由于酶蛋白量的改变。在许多生理条件下,该抗血清已被用于从成年和新生大鼠肝脏经热处理的上清液制剂中沉淀出“苹果酸”酶,这些大鼠在30分钟前已注射了l-[4,5-(³H)]亮氨酸。放射性掺入可免疫沉淀酶的量较低,这使得我们能够定性地得出结论:成年大鼠中酶活性的改变主要源于酶合成速率的变化。断奶大鼠自然发生的或经甲状腺素处理后“苹果酸”酶活性的显著增加同样是由于酶合成速率的显著增加,可能同时伴随着酶降解速率的降低。