Koretsky A P, Wang S, Klein M P, James T L, Weiner M W
Biochemistry. 1986 Jan 14;25(1):77-84. doi: 10.1021/bi00349a012.
31P NMR spectra of rat kidney and heart, in situ, were obtained at 97.2 MHz by using chronically implanted radio-frequency coils. Previous investigators have used magnetization transfer techniques to study phosphorus exchange in perfused kidney and heart. In the current experiments, saturation transfer techniques were used to measure the steady-state rate of exchange between inorganic phosphate (Pi) and the gamma-phosphate of ATP (gamma ATP) in kidney, and between phosphocreatine (PCr) and gamma ATP, catalyzed by creatine kinase, in heart. The rate constant for the exchange detected between Pi and gamma ATP in kidney, presumably catalyzed by oxidative phosphorylation, was 0.12 +/- 0.03 s-1. This corresponds to an ATP synthesis rate of 12 mumol min-1 (g wet weight)-1. Comparison of previously published O2 consumption and Na+ reabsorption rates for the intact kidney with the NMR-derived rate for ATP synthesis gave flux ratios of JATP/JO2 = 1.6-3.3 and JNa+/JATP = 4-10. The rate constants for the creatine kinase reaction, assuming a simple two-site exchange, were found to be 0.57 +/- 0.12 s-1 for the forward direction (PCr----ATP) and 0.50 +/- 0.16 s-1 for the reverse direction (ATP----PCr). The forward rate (0.78 +/- 0.18 intensity unit/s) was significantly larger (p less than 0.05) than the reverse rate (0.50 +/- 0.16 intensity unit/s). This difference between the forward and reverse rates of creatine kinase has been previously noted in the perfused heart. The difference has been attributed to participation of ATP in other reactions.(ABSTRACT TRUNCATED AT 250 WORDS)
采用长期植入的射频线圈,在97.2MHz频率下获取大鼠原位肾脏和心脏的31P核磁共振谱。先前的研究者已使用磁化转移技术研究灌注肾脏和心脏中的磷交换。在当前实验中,采用饱和转移技术测量肾脏中无机磷酸盐(Pi)与ATP的γ-磷酸盐(γATP)之间,以及心脏中由肌酸激酶催化的磷酸肌酸(PCr)与γATP之间的稳态交换速率。肾脏中Pi与γATP之间检测到的交换速率常数(可能由氧化磷酸化催化)为0.12±0.03s-1。这对应于12μmol min-1(湿重g)-1的ATP合成速率。将先前发表的完整肾脏的氧气消耗率和钠重吸收率与核磁共振得出的ATP合成速率进行比较,得出通量比JATP/JO2 = 1.6 - 3.3和JNa+/JATP = 4 - 10。假设为简单的两点交换,肌酸激酶反应的速率常数在正向(PCr→ATP)为0.57±0.12s-1,在反向(ATP→PCr)为0.50±0.16s-1。正向速率(0.78±0.18强度单位/秒)显著大于(p<0.05)反向速率(0.50±0.16强度单位/秒)。肌酸激酶正向和反向速率之间的这种差异先前在灌注心脏中已被注意到。这种差异归因于ATP参与其他反应。(摘要截于250字)