Vrbjar N, Dzurba A, Ziegelhöffer A
Department of Biochemistry, Slovak Academy of Sciences, Bratislava.
Mol Cell Biochem. 1995;147(1-2):99-103. doi: 10.1007/BF00944789.
To elucidate the effect of global ischemia on the energy utilizing processes, regarding the molecular principles, the kinetic and thermodynamic properties of the sarcolemmal ATPases were investigated in the rat heart. The activation energy for hydrolysis of ATP during ischemia was higher when the reaction was catalyzed by Ca-ATPase or Mg-ATPase. For the Na,K-ATPase reaction, no changes in the activation energy were observed. With respect to the enzyme kinetics, ischemia in a time-dependent manner induced important alterations in KM and Vmax values of Na,K-ATPase, Ca-ATPase and Mg-ATPase. The Vmax value decreased significantly already after 15 min of ischemia, and it also remained low after 30, 45 and 60 min for all 3 enzymes. The significant diminution of KM values occurred later in the 30th min for Ca-ATPase, in the 45th min for Na,K-ATPase. The observed drop in KM indicates the increase in the affinity of the enzymes to substrate, suggesting thus the adaptation to ischemic conditions on the molecular level. This effect could be attributed to some conformational changes of the protein molecule in the vicinity of the ATP-binding site developing after longer duration of ischemia.
为了阐明整体缺血对能量利用过程的影响,从分子原理方面,对大鼠心脏肌膜ATP酶的动力学和热力学性质进行了研究。当由钙ATP酶或镁ATP酶催化反应时,缺血期间ATP水解的活化能更高。对于钠钾ATP酶反应,未观察到活化能的变化。就酶动力学而言,缺血以时间依赖性方式引起钠钾ATP酶、钙ATP酶和镁ATP酶的米氏常数(KM)和最大反应速度(Vmax)值发生重要改变。缺血15分钟后,Vmax值就显著降低,并且在30、45和60分钟后,这三种酶的Vmax值也都保持在较低水平。钙ATP酶的KM值在第30分钟时显著降低,钠钾ATP酶的KM值在第45分钟时显著降低。观察到的KM值下降表明酶与底物的亲和力增加,从而表明在分子水平上对缺血条件的适应。这种效应可能归因于缺血持续较长时间后在ATP结合位点附近蛋白质分子发生的一些构象变化。