Kuo D J, Dikdan G, Jordan F
J Biol Chem. 1986 Mar 5;261(7):3316-9.
A novel purification method was developed for brewers' yeast pyruvate decarboxylase (EC 4.1.1.1) that for the first time resolved the enzyme into two isozymes on DEAE-Sephadex chromatography. The isozymes were found to be distinct according to sodium dodecyl sulfate polyacrylamide gel electrophoresis: the first one to be eluted gave rise to one band, the second to two bands. The isozymes were virtually the same so far as specific activity, KM, inhibition kinetics and irreversible binding properties by the mechanism-based inhibitor (E)-4-(4-chlorophenyl)-2-oxo-3-butenoic acid are concerned. This finding resolves a longstanding controversy concerning the quaternary structure of this enzyme.
开发了一种用于啤酒酵母丙酮酸脱羧酶（EC 4.1.1.1）的新型纯化方法，该方法首次在DEAE-葡聚糖凝胶色谱上将该酶分离为两种同工酶。根据十二烷基硫酸钠聚丙烯酰胺凝胶电泳发现，这些同工酶是不同的：第一个被洗脱的产生一条带，第二个产生两条带。就比活性、米氏常数（KM）、抑制动力学以及基于机制的抑制剂（E）-4-（4-氯苯基）-2-氧代-3-丁烯酸的不可逆结合特性而言，这些同工酶几乎相同。这一发现解决了关于该酶四级结构的长期争议。
