Price N C, Stevens E
Biochem J. 1983 Mar 1;209(3):763-70. doi: 10.1042/bj2090763.
The refolding of rabbit muscle pyruvate kinase after denaturation by guanidine hydrochloride was studied. On dilution of the denaturing agent, enzyme activity is only partially regained. The extent of regain of activity is dependent on protein concentration, showing a marked decrease at higher concentrations. The failure to regain complete activity appears to be related to the formation of inactive aggregates, which can be separated from active enzyme by gel filtration. Insoluble aggregates can be partially re-activated after solubilization in guanidine hydrochloride. Changes in the circular-dichroism and fluorescence spectra during refolding suggest that a partially folded, inactive species is formed rapidly; this differs from native enzyme in being more susceptible to proteolysis by trypsin.
研究了盐酸胍变性后兔肌肉丙酮酸激酶的复性。在稀释变性剂时，酶活性仅部分恢复。活性恢复的程度取决于蛋白质浓度，在较高浓度时显著降低。未能完全恢复活性似乎与无活性聚集体的形成有关，可通过凝胶过滤将其与活性酶分离。不溶性聚集体在盐酸胍中溶解后可部分重新激活。复性过程中圆二色性和荧光光谱的变化表明，一种部分折叠的无活性物种迅速形成；这与天然酶不同，它更容易被胰蛋白酶水解。
