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类固醇和氯化铵酸中毒对大鼠肾皮质磷酸烯醇丙酮酸羧激酶的影响。I. 诱导过程的分化及酶活性的表征

The effect of steroids and ammonium chloride acidosis on phosphoenolpyruvate carboxykinase in rat kidney cortex. I. Differentiation of the inductive process and characterization of enzyme activities.

作者信息

Longshaw I D, Pogson C I

出版信息

J Clin Invest. 1972 Sep;51(9):2277-83. doi: 10.1172/JCI107037.

Abstract

The behaviour of rat kidney cortex phosphoenolpyruvate carboxykinase has been investigated under conditions of triamcinolone administration and ammonium chloride acidosis. The concentration of phosphoenolpyruvate carboxykinase as measured by enzyme activity and immunotitration was elevated under both conditions. The mechanism of induction is different in the two cases. At doses which produce maximum stimulation, the effects of steroid and ammonium chloride were additive; only the increment in enzyme activity produced by steroid was blocked by actinomycin D. PHOSPHOENOLPYRUVATE CARBOXYKINASE ACTIVITIES IN ALL CONDITIONS INVESTIGATED SHOW SIMILAR BEHAVIOR IN DILUTE EXTRACTS: these experiments involved antibody titration, stability studies, and molecular weight determinations on sucrose gradients. The molecular weight of phosphoenolpyruvate carboxykinase was also studied in undiluted extracts prepared by high-speed centrifugation; values were determined from sedimentation data obtained with a moving-partition cell as described by Yphantis and Waugh. Under these conditions, the apparent molecular weight of phosphoenolpyruvate carboxykinase was increased from 83,000 to 128,000 by ammonium chloride acidosis. These results are discussed and a hypothesis regarding the mechanism of phosphoenolpyruvate carboxykinase regulation in kidney cortex is presented.

摘要

在给予曲安西龙和氯化铵酸中毒的条件下,对大鼠肾皮质磷酸烯醇丙酮酸羧激酶的行为进行了研究。在这两种情况下,通过酶活性和免疫滴定法测得的磷酸烯醇丙酮酸羧激酶浓度均升高。两种情况下诱导机制不同。在产生最大刺激的剂量下,类固醇和氯化铵的作用是相加的;只有类固醇产生的酶活性增加被放线菌素D阻断。在所研究的所有条件下,磷酸烯醇丙酮酸羧激酶活性在稀释提取物中表现出相似行为:这些实验涉及抗体滴定、稳定性研究以及在蔗糖梯度上的分子量测定。还通过高速离心制备的未稀释提取物研究了磷酸烯醇丙酮酸羧激酶的分子量;如Yphantis和Waugh所述,根据用移动隔板池获得的沉降数据确定数值。在这些条件下,氯化铵酸中毒使磷酸烯醇丙酮酸羧激酶的表观分子量从83,000增加到128,000。对这些结果进行了讨论,并提出了关于肾皮质中磷酸烯醇丙酮酸羧激酶调节机制的假说。

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