Rose bengal as a tool in studying the ligand binding of phosphorylase b.

L Trón, J Matkó
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引用次数: 0

Abstract

The interaction of rose bengal (RB) with rabbit skeletal muscle phosphorylase b (1,4-alpha-D glucan: orthophosphate alpha-glucosyl-transferase, E.C. 2.4.1.1.) was studied by kinetic and absorption photometric methods. RB inhibited the phosphorylase b activity. Inhibition was strictly competitive with respect to substrate G-1-P and activator AMP with inhibition constants 2 x 10(-6) M and 2.2. x 10(-7) M, respectively. The association of the dye with the enzyme elicited a red shift in the spectrum of RB indicating an apolar binding site. According to difference absorption measurements, the enzyme binds two dye molecules per dimer in the presence and absence of both G-1-P and AMP. Binding constants determined from photometric titrations are consistent with those obtained from kinetic measurements. The present findings allow to carry out detailed kinetic investigations on the activator AMP and substrate G-1-P binding of phosphorylase b.

以玫瑰为工具研究磷酸化酶b的配体结合。
采用动力学和吸收光度法研究了玫瑰花(RB)与兔骨骼肌磷酸化酶b (1,4- α - d葡聚糖:正磷酸α -葡萄糖基转移酶,E.C. 2.4.1.1)的相互作用。RB抑制磷酸化酶b活性。底物G-1-P和活化剂AMP的抑制常数分别为2 × 10(-6) M和2.2。x 10(-7) M。染料与酶的结合引起RB光谱的红移,表明极性结合位点。根据差异吸收测量,该酶在G-1-P和AMP存在和不存在的情况下,每个二聚体结合两个染料分子。光度滴定测定的结合常数与动力学测量结果一致。本研究结果允许对磷酸化酶b的激活剂AMP和底物G-1-P结合进行详细的动力学研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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