Schulz J, Wilhelm G, Lorenz G, Hofmann E
Acta Biol Med Ger. 1975;34(8):1321-32.
Pyruvate kinase from ascites tumour cells can be eluted from phosphocellulose by very low concentrations of phosphoenolpyruvate, fructose 1,6-bisphosphate, adenosine 5'-diphosphate and pyrophosphate, respectively. The appropriate limiting conditions for "facilitated desorption" of the enzyme from phosphocellulose by these ligands have been elaborated for achieving maximum selectivity and recovery in the process of its purification. This method has been designated as "affinity elution chromatography" owing to the specific interactions between a ligand as a constituent of the eluting medium with the adsorbed enzyme, which causes its selective desorption from the ion-exchanger. Affinity elution with phosphoenolpyruvate has been found to be very effective for preparation of the M-types of pyruvate kinase. A specific activity of 420 for an almost homogeneous preparation of pyruvate kinase from ascites tumour cells has maximally been obtained.
腹水肿瘤细胞中的丙酮酸激酶可以分别用极低浓度的磷酸烯醇丙酮酸、果糖1,6 -二磷酸、腺苷5'-二磷酸和焦磷酸从磷酸纤维素上洗脱下来。已经阐述了这些配体从磷酸纤维素上“促进解吸”该酶的适当极限条件，以便在其纯化过程中实现最大的选择性和回收率。由于作为洗脱介质成分的配体与吸附的酶之间存在特异性相互作用，这种相互作用导致酶从离子交换剂上选择性解吸，所以该方法被称为“亲和洗脱色谱法”。已发现用磷酸烯醇丙酮酸进行亲和洗脱对于制备丙酮酸激酶的M型非常有效。从腹水肿瘤细胞中获得的几乎纯的丙酮酸激酶制剂的比活性最高可达420。
