Barbotin J N, Thomasset B
Biochim Biophys Acta. 1979 Sep 12;570(1):11-21. doi: 10.1016/0005-2744(79)90196-7.
Active soluble cross-linked L-glutamate dehydrogenase (L-glutamate: NAD(P)+ oxidoreductase (deaminating), EC 1.4.1.3) albumin polymers were produced. Electron microscopic studies and kinetic properties were studied with the polymer in solution and compared with previous published data about the enzyme immobilized inside proteic films (Barbotin, J.N. and Breuil, M. (1978) Biochim. Biophys. Acta 525, 18--27). The glutaraldehyde effect on activity yield, ADP and beta-NAD+ protection, stability and pH rate profile were studied and discussed. Apparent Michaelis constants were determined with soluble polymers produced with or without ADP during the grafting process. Experiments were performed on the regulatory properties of immobilized glutamate dehydrogeanse showing the decrease of ADP activation and GTP inhibition as compared to the free form. In other respects, electron microscopy observations showed morphological differences between the two populations of soluble polymers produced in presence of ADP, obtained after gel filtration on Sepharose 6B. Linear aggregates of high molecular weight and classical soluble polymers were obtained. Similar Km values and regulatory properties were exhibited by the two forms, demonstrating the absence of interdependence between the allosteric control and the polymerization of enzyme monomers.
制备了活性可溶性交联L-谷氨酸脱氢酶（L-谷氨酸：NAD(P)+氧化还原酶（脱氨基），EC 1.4.1.3）白蛋白聚合物。对溶液中的聚合物进行了电子显微镜研究和动力学性质研究，并与先前发表的关于固定在蛋白质膜内的酶的数据进行了比较（Barbotin, J.N.和Breuil, M.（1978年）《生物化学与生物物理学报》525, 18 - 27）。研究并讨论了戊二醛对活性产率、ADP和β-NAD+保护、稳定性和pH速率曲线的影响。在接枝过程中，用有或无ADP产生的可溶性聚合物测定了表观米氏常数。对固定化谷氨酸脱氢酶的调节特性进行了实验，结果表明与游离形式相比，ADP激活和GTP抑制作用降低。在其他方面，电子显微镜观察显示，在Sepharose 6B上凝胶过滤后得到的两种在ADP存在下产生的可溶性聚合物群体之间存在形态差异。得到了高分子量的线性聚集体和经典的可溶性聚合物。两种形式表现出相似的Km值和调节特性，表明别构控制和酶单体聚合之间不存在相互依赖性。
