Adenosine attenuates C-terminal but not N-terminal proteolysis of cTnI during cardioplegic arrest.
BACKGROUND
Specific site proteolysis and loss of troponin I (TnI) during myocardial ischemic events can contribute to myocardial dysfunction. Adenosine supplementation of cardioplegic solutions results in improved functional preservation of the heart. We investigated the effect of adenosine on N-terminal and C-terminal proteolysis of TnI in the heart.


METHODS
Hearts from male Sprague-Dawley rats were isolated and perfused at a constant pressure. Cardioplegic arrest (St. Thomas #2 +/- 100 microm adenosine) was induced and hearts frozen at various times during the arrest. Antibodies directed against specific regions of TnI were used to visualize TnI in whole heart homogentates, as well as from cellular fractions, using western blot analysis.


RESULTS
Cardioplegic arrest alone resulted in early N-terminal proteolysis of TnI, followed by later loss of sequences from the C-terminal end of the molecule. In addition, secondary protein bands that were immunoreactive to amino acid sequences centrally located on the TnI molecule were observed. There was also evidence of dissociation of TnI from the other myofibrillar proteins. The supplementation of cardioplegic solution with adenosine significantly attenuated the late C-terminal proteolytic degradation of TnI and its apparent dissociation from myofibrils proteins but had no effect on the early N-terminal proteolysis associated with cardioplegic arrest.


CONCLUSIONS
These data may provide an explanation for partial protection against postarrest myocardial dysfunction provided by adenosine.
Law
William R
WR
West Side Veterans Administration Medical Center, Research Service, Chicago, Illinois, USA. wrlaw@uic.edu
Ross
James D
JD
Jonjev
Zivojin S
ZS
D000241:Adenosine / Q000494:pharmacology*; D000818:Animals; D006324:Heart Arrest, Induced; D008297:Male; D009206:Myocardium / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013599:Systole / Q000187:drug effects; D019210:Troponin I / Q000032:analysis / Q000378:metabolism*
D019210:Troponin I; D000241:Adenosine