Is creatine kinase a target for AMP-activated protein kinase in the heart?
By phosphorylating target proteins, AMP-activated protein kinase (AMPK) inhibits ATP-utilizing proteins and activates ATP-synthesizing proteins, thereby increasing ATP synthesis under conditions such as hypoxia and ischemia. It has been proposed that AMPK also phosphorylates and inhibits creatine kinase (CK), the enzyme which catalyzes the reversible transfer of a phosphoryl group between creatine and ADP. Here, we examine the hypothesis that AMPK inactivates CK activity under three conditions where [AMP] and AMP-dependent AMPK velocity increase: increased workload both in the isolated rat heart and in the living rat, hypoxia in the living rat heart and low-flow ischemia in the isolated red blood cell perfused rat heart. For the experiments varying workload in the isolated rat heart (both ejecting and isovolumic models), we also changed oxidizable substrate available to the isolated heart in order to vary the [AMP]/[ATP]. CK reaction velocity in the intact rat heart was directly measured using (31)P magnetization transfer. The metabolically active AMP and ATP pools were determined from (31)P NMR measurements and we calculate AMP-dependent AMPK velocity from the Michaelis-Menten relationship. We found that under normoxic conditions where [AMP] and AMPK velocity increase, the linear relationship between CK and AMPK velocities is positive, not inverse. Under conditions of low pO(2) (hypoxia and low-flow ischemia), CK velocity fell 2-4-fold while the increase in AMP-activated AMPK activity was modest. This analysis illustrates the complex nature of AMPK regulation in the heart.
Ingwall
Joanne S
JS
Division of Cardiovascular Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA. jingwall@rics.bwh.harvard.edu
D000263:Adenylate Kinase / Q000378:metabolism*; D000818:Animals; D003402:Creatine Kinase / Q000378:metabolism*; D004734:Energy Metabolism; D004789:Enzyme Activation; D000860:Hypoxia; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D017202:Myocardial Ischemia / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism*; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley
D003402:Creatine Kinase; D000263:Adenylate Kinase