Vas M, Batke J
Biochim Biophys Acta. 1981 Aug 13;660(2):193-8. doi: 10.1016/0005-2744(81)90159-5.
The possibility of a functional complex formation between glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12) and 3-phosphoglycerate kinase (EC. 2.7.2.3), enzymes catalysing two consecutive reactions in glycolysis has been investigated. Kinetic analysis of the coupled enzymatic reaction did not reveal any kinetic sign of the assumed interaction up to 4 X 10(-6) M kinase and 10(-4) M dehydrogenase. Fluorescence anisotrophy of 10(-7) M or 2 X 10(-5) M glyceraldehyde-3-phosphate dehydrogenase labeled with fluorescein isothiocynate did not change in the presence of non-labeled 3-phosphoglycerate kinase (up to 4 X 10(-5) M). The frontal gel chromatographic analysis of a mixture of the two enzymes (10(-4) M dehydrogenase) could not reveal any molecular species with the kinase activity having a molecular weight higher than that of 3-phosphoglycerate kinase. Both types of physicochemical measurements were also performed in the presence of substrates of the kinase and gave the same results. The data seem to invalidate the hypothesis that there is a complex between purified pig muscle glyceraldehyde-3-phosphate dehydrogenase and 3-phosphoglycerate kinase.
对甘油醛-3-磷酸脱氢酶（EC 1.2.1.12）和3-磷酸甘油酸激酶（EC. 2.7.2.3）之间形成功能复合物的可能性进行了研究，这两种酶催化糖酵解中的两个连续反应。对耦合酶促反应的动力学分析表明，在激酶浓度高达4×10⁻⁶ M和脱氢酶浓度高达10⁻⁴ M时，未发现假定相互作用的任何动力学迹象。用异硫氰酸荧光素标记的10⁻⁷ M或2×10⁻⁵ M甘油醛-3-磷酸脱氢酶的荧光各向异性，在存在未标记的3-磷酸甘油酸激酶（浓度高达4×10⁻⁵ M）时没有变化。对两种酶的混合物（10⁻⁴ M脱氢酶）进行的前沿凝胶色谱分析，未发现任何具有激酶活性且分子量高于3-磷酸甘油酸激酶的分子种类。这两种物理化学测量也在激酶底物存在的情况下进行，结果相同。这些数据似乎否定了纯化的猪肌肉甘油醛-3-磷酸脱氢酶和3-磷酸甘油酸激酶之间存在复合物的假设。
