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不同年龄大鼠肝脏匀浆中的γ-谷氨酰转肽酶:酶动力学以及米氏常数(Km)和最大反应速度(Vmax)的年龄变化过程

gamma-Glutamyltranspeptidase in liver homogenates of rats of different ages: enzyme kinetics and age course of Km and Vmax.

作者信息

Kretzschmar M, Klinger W

出版信息

Z Versuchstierkd. 1989;32(1):41-7.

PMID:2565636
Abstract

Optimum incubation conditions for the determination of gamma-glutamyltranspeptidase with liver homogenates from rats of different ages have been determined: about 1 mg protein/assay, 4.4 mM L-gamma-glutamyl-p-nitroanilide, 40 mM glycylglycine, 9.6 mM MgCl2 in 0.1 M Tris-HCl buffer, pH 8.0; final volume 1.3 ml. The enzyme activity is very high at birth, the increase before birth is not continuous. After birth the enzyme activity decreases quickly over the first postnatal days. The ENDRENYI-KWONG-plot was linear with homogenates from newborn and 3-day-old animals. For all other age groups the graphs showed an angle. Statistical analysis carried out that a two-enzyme model plots the experimental data only insignificantly better than a one-enzyme model. It may be concluded that there are different GGT isozymes which show different affinities towards the substrate and which show different developmental patterns.

摘要

已确定用不同年龄大鼠的肝脏匀浆测定γ-谷氨酰转肽酶的最佳孵育条件:每次测定约1mg蛋白质、4.4mM L-γ-谷氨酰对硝基苯胺、40mM甘氨酰甘氨酸、9.6mM氯化镁,溶于0.1M Tris-HCl缓冲液(pH 8.0)中;终体积1.3ml。出生时酶活性非常高,出生前的增加不连续。出生后,酶活性在出生后的头几天迅速下降。对于新生动物和3日龄动物的匀浆,恩德里尼-邝氏图呈线性。对于所有其他年龄组,图表显示有一个角度。统计分析表明,双酶模型绘制实验数据仅比单酶模型略好。可以得出结论,存在不同的γ-谷氨酰转肽酶同工酶,它们对底物表现出不同的亲和力,并且表现出不同的发育模式。

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