Cardiac proteasome dysfunction during cold ischemic storage and reperfusion in a murine heart transplantation model.
Recent observations suggest that the ubiquitin-proteasome system (UPS) contributes to the pathophysiology of myocardial ischemia-reperfusion injury. Since its regulation during cold ischemia-reperfusion is unknown, we evaluated the cardiac UPS in a model of heart transplantation in mice. Cardiac ubiquitylation rates and ubiquitin-protein conjugates increased after 3h of cold ischemia (CI) and normalized post-transplant. 20S proteasome content and proteasome peptidase activities were unchanged after CI. 4h/24h post-transplant 20S proteasome concentrations decreased and chymotryptic-like but not tryptic-like proteasome peptidase activity was inactivated. Epoxomicin sensitivity of the proteasome increased 5.7-fold during CI and normalized 4h/24h post-transplant. This was accompanied by the disappearance of a 13.5 kDa-ubiquitin-conjugate during CI that could be attenuated by addition of epoxomicin to the preservation fluid. We conclude that substrate specificity of the proteasome changes during cold ischemia and that proteasome inhibition preserves the physiological ubiquitin-protein conjugate pool during organ preservation. Reduced proteasome activity during reperfusion is caused by a decrease in proteasome content and enzyme inhibition.
Majetschak
Matthias
M
DeWitt Daughtry Family Department of Surgery, Divisions of Trauma and Surgical Critical Care - Trauma Research and Cardiothoracic Surgery, University of Miami Miller School of Medicine, 1800 NW 10th Avenue, Miami, FL 33136, USA.
Patel
Mayur B
MB
Sorell
Luis T
LT
Liotta
Carlos
C
Li
Sen
S
Pham
Si M
SM
D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D016027:Heart Transplantation / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D023421:Models, Animal; D009206:Myocardium / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D015424:Reperfusion / Q000379:methods*
D020543:Proteome; D046988:Proteasome Endopeptidase Complex