Effects of peroxynitrite on isolated cardiac trabeculae: selective impact on myofibrillar energetic controllers.
Formation of peroxynitrite and cardiac protein nitration have been implicated in multiple cardiac disease states, but their contributions to disease initiation remain undefined. We have previously observed nitration of myofibrillar regions of cardiac myocytes in several experimental and clinical settings of cardiac myocyte dysfunction and postulated that oxidative insult to key components of the contractile apparatus may be initiating events. Here we tested the hypothesis that peroxynitrite alters myofibrillar contractile function, and investigated a mechanistic role for nitration in this process. Isolated rat ventricular trabeculae were exposed to physiologically relevant concentrations of peroxynitrite and ATP-dependent contractile responses were measured. Maximal trabecular force generation was significantly impaired following 300 nM peroxynitrite exposures. Several myofibrillar proteins demonstrated increased tyrosine nitration, the most significant increases occurred in the myosin heavy chain and the myofibrillar isoform of creatine kinase. Additional functional experiments were conducted using phosphocreatine (high energy phosphate substrate for myofibrillar creatine kinase) as the primary energy substrate. Myofibrillar creatine kinase-dependent force generation was impaired at peroxynitrite concentrations as low as 50 nM, suggesting potent inactivation of the enzyme. Extent of tyrosine nitration of myofibrillar creatine kinase was negatively correlated to myofibrillar creatine kinase-dependent force generation. These data demonstrate that the cardiac contractile apparatus is highly sensitive to peroxynitrite, and that MM-CK may be a uniquely vulnerable target.
Mihm
Michael J
MJ
Center for Developmental Pharmacology and Toxicology, Columbus Children's Research Institute, 700 Children's Drive, Columbus, OH 43205, USA.
Yu
Fushun
F
Reiser
Peter J
PJ
Bauer
John Anthony
JA
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D003402:Creatine Kinase / Q000378:metabolism; D004734:Energy Metabolism / Q000187:drug effects*; D006321:Heart / Q000033:anatomy & histology / Q000187:drug effects / Q000502:physiology; D008297:Male; D009200:Myocardial Contraction / Q000187:drug effects; D009206:Myocardium / Q000201:enzymology / Q000378:metabolism*; D009566:Nitrates / Q000378:metabolism; D030421:Peroxynitrous Acid / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014443:Tyrosine / Q000378:metabolism
D009566:Nitrates; D030421:Peroxynitrous Acid; D014443:Tyrosine; D003402:Creatine Kinase; D000251:Adenosine Triphosphatases