Viglio S, Valentini G, Chiarelli L, Zanaboni G, Cetta G, Iadarola P
Dipartimento di Biochimica A. Castellani, Università di Pavia, Italy.
Electrophoresis. 1999 Sep;20(12):2400-6. doi: 10.1002/(SICI)1522-2683(19990801)20:12<2400::AID-ELPS2400>3.0.CO;2-T.
Micellar electrokinetic chromatography (MEKC) has been utilized as an analytical method to perform investigations on limited proteolysis of proteins. To this purpose partial proteolysis experiments with a series of proteinases were performed, utilizing as model protein pyruvate kinase (PK) from Escherichia coli, an enzyme that is regulated allosterically by fructose 1,6-bisphosphate (FBP). Data obtained with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and MEKC were compared; the profiles generated by submitting digests of PK treated with different proteinases in the presence and absence of FBP to electrophoretic analysis provided a useful adjunct for a better understanding of the effects of the allosteric activator on the conformation of the model enzyme. MEKC was also found to be a convenient technique for determining the kinetics of substrate proteolysis.
胶束电动色谱法（MEKC）已被用作一种分析方法，用于对蛋白质的有限蛋白酶解进行研究。为此，使用一系列蛋白酶进行了部分蛋白酶解实验，以大肠杆菌的丙酮酸激酶（PK）作为模型蛋白，该酶受1,6-二磷酸果糖（FBP）变构调节。比较了通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和MEKC获得的数据；在有和没有FBP的情况下，将用不同蛋白酶处理的PK消化物进行电泳分析所产生的图谱，为更好地理解变构激活剂对模型酶构象的影响提供了有用的辅助手段。还发现MEKC是一种用于确定底物蛋白酶解动力学的便捷技术。
