Harrigan P J, Trentham D R
Biochem J. 1974 Nov;143(2):353-63. doi: 10.1042/bj1430353.
The kinetics of the acylation of d-glyceraldehyde 3-phosphate dehydrogenase from pig muscle by 1,3-diphosphoglycerate in the presence of NAD(+) has been analysed by using the relaxation temperature-jump method. At pH7.2 and 8 degrees C the rate of acylation of the NAD(+)-bound (or holo-) enzyme was 3.3x10(5)m(-1).s(-1) and the rate of phosphorolysis, the reverse reaction, was 7.5x10(3)m(-1).s(-1). After a temperature-jump perturbation the equilibrium of NAD(+) binding to the acyl-enzyme was re-established more rapidly than that of the acylation. The rate of phosphorolysis of the apoacylenzyme from sturgeon muscle and of aldehyde release from the d-glyceraldehyde 3-phosphate-apoenzyme complex were </=40m(-1).s(-1) and </=12s(-1) respectively at pH8.0 and 22 degrees C, which means that both processes are too slow to contribute significantly to the reaction pathway of the reversible NAD(+)-linked oxidative phosphorylation of d-glyceraldehyde 3-phosphate. Phosphorolysis of both acyl-apoenzyme and acyl-holoenzyme was first-order in P(i) up to 100mm-P(i) and more. PO(4) (3-) could be the reactive species of the phosphorolysis of the acyl-holoenzyme, in which case phosphorolysis is a diffusion-controlled reaction, although other kinetically indistinguishable rate equations for the reaction are possible.
利用弛豫温度跃升法分析了在NAD(+)存在下，1,3 -二磷酸甘油酸对猪肌肉中d -甘油醛3 -磷酸脱氢酶的酰化动力学。在pH7.2和8℃时，与NAD(+)结合的（或全酶）酶的酰化速率为3.3×10(5)m(-1).s(-1)，而磷酸解（逆反应）速率为7.5×10(3)m(-1).s(-1)。温度跃升扰动后，NAD(+)与酰基酶结合的平衡比酰化平衡更快地重新建立。在pH8.0和22℃时，鲟鱼肌肉中脱辅基酰基酶的磷酸解速率和d -甘油醛3 -磷酸 - 脱辅基酶复合物中醛释放速率分别≤40m(-1).s(-1)和≤12s(-1)，这意味着这两个过程都太慢，对d -甘油醛3 -磷酸可逆的NAD(+)连接的氧化磷酸化反应途径贡献不大。酰基 - 脱辅基酶和酰基 - 全酶的磷酸解在磷酸根离子浓度高达100mmol/L及更高时对磷酸根离子呈一级反应。磷酸根离子（PO(4)(3-)）可能是酰基 - 全酶磷酸解的反应活性物种，在这种情况下，磷酸解是一个扩散控制反应，尽管该反应其他动力学上无法区分的速率方程也是可能的。
