牛脑M1型丙酮酸激酶的纯化及性质

Purification and properties of pyruvate kinase type M1 from bovine brain.

Terlecki G

Department of Biochemistry, Medical School of Wrocław, Poland.

Int J Biochem. 1989;21(9):1053-60. doi: 10.1016/0020-711x(89)90240-1.

Abstract

Pyruvate kinase type M1 was purified from bovine brain about 241-fold with 38% yield. 2. Specific activity of the enzyme was above 217 U/mg of protein (25 degrees C), relative mol. wt of the subunit--57,000 (+/- 2000) and pH optimum--6.8-7.2. 3. The enzyme shoved hyperbolic kinetics with Km value for PEP of 0.04 mM and for ADP of 0.3 mM. 4. Inorganic phosphate and ATP at concentrations below 4 mM showed activating effect, 1-phenylalanine and ATP above 6 mM--an inhibiting effect on the enzyme. 5. Inhibition by 1-phenylalanine was prevented by fructose-1,6-bisphosphate.

摘要

丙酮酸激酶M1型从牛脑中纯化出来，纯化倍数约为241倍，产率为38%。2. 该酶的比活性在25℃时高于217 U/mg蛋白质，亚基的相对分子量为57,000（±2000），最适pH为6.8 - 7.2。3. 该酶呈现双曲线动力学，对磷酸烯醇式丙酮酸（PEP）的Km值为0.04 mM，对二磷酸腺苷（ADP）的Km值为0.3 mM。4. 浓度低于4 mM的无机磷酸盐和ATP表现出激活作用，浓度高于6 mM的1 - 苯丙氨酸和ATP对该酶有抑制作用。5. 1 - 苯丙氨酸的抑制作用可被1,6 - 二磷酸果糖阻止。