Solov'eva G A, Sundukov S Iu, Filippova G N
Vopr Med Khim. 1990 May-Jun;36(3):88-91.
A turbidimetric procedure is described, which involves the monitoring of changes in glycogen turbidity at wavelengths above 300 nm and continuous recording of rabbit skeletal muscle synthase activity. The recalculation coefficients were found to be equal to 1.69 +/- 0.08 mM UDP per unit of optical density at 360 nm and to 2.03 +/- 0.01 mM UDP per unit of optical density at 400 nm. The procedure allows a kinetic analysis of the enzyme within a broad range of concentrations and under various conditions. The glycogen synthase activity did not depend on the buffer capacity when 10-100 mM Tris-HCL buffer, pN 7.8, was used. The rate of the enzymatic reaction was correlated with the enzyme concentration within the range of 5 to 50 micrograms/ml. The curve for glycogen synthase saturation with UDPG is described by the Michaelis-Menten equation, when either 0.04-0.08 mM glucose-6-phosphate for for the D-form were used in mixtures containing 5 mM MgCl2 for the D-form were used in mixtures containing 5 mM MgCl2 and 10 mM Na2SO4. The turbidimetric and spectrophotometric procedures yielded similar results.
本文描述了一种比浊法,该方法涉及监测波长大于300nm时糖原浊度的变化,并连续记录兔骨骼肌合酶活性。经重新计算,在360nm处每单位光密度的系数等于1.69±0.08mM UDP,在400nm处每单位光密度的系数等于2.03±0.01mM UDP。该方法可以在很宽的浓度范围和各种条件下对该酶进行动力学分析。当使用10 - 100mM Tris-HCL缓冲液(pH 7.8)时,糖原合酶活性不依赖于缓冲容量。酶促反应速率与5至50微克/毫升范围内的酶浓度相关。当在含有5mM MgCl2和10mM Na2SO4的混合物中,对于D型使用0.04 - 0.08mM葡萄糖-6-磷酸时,UDPG对糖原合酶的饱和曲线可用米氏方程描述。比浊法和分光光度法得出了相似的结果。