Gerschitz J, Rudolph R, Jaenicke R
Eur J Biochem. 1978 Jul 3;87(3):591-9. doi: 10.1111/j.1432-1033.1978.tb12411.x.
Horse-liver alcohol dehydrogenase has been dissociated and denatured by 6 M guanidinium hydrochloride. Removal of the denaturant under optimum conditions of the solvent leads to partial reactivation. The concentrations of the enzyme, as well as the coenzyme (NAD+), and Zn2+, affect the reactivation significantly, since high concentrations promote the formation of inactive aggregation products. Analyzing the kinetics of reactivation and reassociation, conditions far from equilibrium of dissociation-association provide maximum yields (approximately 70%). The sigmoidal kinetic traces suggest a superposition of first-order transconformation and second-order association reactions; the latter are corroborated by the concentration dependence of the reactivation reaction. The coenzyme, NAD+, has no influence on the kinetics of reactivation. Addition of Zn2+ leads to a significant decrease of the rate and yield of reactivation. The process of renaturation, as reflected by the regain of native fluorescence shows complex kinetics: rapid relaxations are followed by slower first-order and second-order processes which parallel reactivation.
马肝醇脱氢酶已被6M盐酸胍解离和变性。在溶剂的最佳条件下去除变性剂会导致部分重新活化。酶的浓度以及辅酶（NAD +）和Zn2 +对重新活化有显著影响，因为高浓度会促进无活性聚集产物的形成。分析重新活化和重新缔合的动力学，远离解离 - 缔合平衡的条件可提供最大产率（约70%）。S形动力学曲线表明一级转构象和二级缔合反应的叠加；后者通过重新活化反应的浓度依赖性得到证实。辅酶NAD +对重新活化的动力学没有影响。添加Zn2 +会导致重新活化的速率和产率显著降低。如天然荧光恢复所反映的复性过程显示出复杂的动力学：快速弛豫之后是较慢的一级和二级过程，这些过程与重新活化平行。
