Phosphorylation and translocation of heat shock protein 27 and alphaB-crystallin in human myocardium after cardioplegia and cardiopulmonary bypass.
OBJECTIVE
Cardiac surgery using cardioplegia and cardiopulmonary bypass subjects myocardium to hypothermic reversible ischemic injury that can impair cardiac function. Research in animal and cell models demonstrates that acute myocardial ischemia/reperfusion injury causes phosphorylation of heat shock protein 27 and alphaB-crystallin. Phosphorylation of heat shock protein 27 and alphaB-crystallin is implicated in the regulation of both beneficial and detrimental responses to ischemic injury. The phosphorylation status of these proteins in human myocardium after ischemic insults associated with cardioplegia and cardiopulmonary bypass is unknown.


METHODS
Right atrial appendage and chest wall skeletal muscle samples were collected from patients before and after cardioplegia and cardiopulmonary bypass. Cardioplegia and cardiopulmonary bypass-induced changes in phosphorylation and localization of heat shock protein 27 and alphaB-crystallin were determined using immunoblot and confocal microscopy with total and phospho-specific antibodies.


RESULTS
Cardioplegia and cardiopulmonary bypass increased the phosphorylation of heat shock protein 27 on serine 15, 78, and 82, and alphaB-crystallin on serine 59 and 45, but not serine 19. The majority of heat shock protein 27 and alphaB-crystallin localized to I-bands of cardiac myofilaments and shifted to a detergent insoluble fraction after cardioplegia and cardiopulmonary bypass. Cardioplegia and cardiopulmonary bypass-induced phosphorylation of specific heat shock protein 27 and alphaB-crystallin residues were associated with additional subcellular locations. Increases in phosphorylation of heat shock protein 27 and alphaB-crystallin were negatively correlated with cardiac function after surgery.


CONCLUSIONS
Cardiac surgery using cardioplegia and cardiopulmonary bypass is associated with phosphorylation and myofilament translocation of heat shock protein 27 and alphaB-crystallin in human myocardium. Phosphorylation of specific heat shock protein 27 and alphaB-crystallin serine residues is associated with distinct localization. Understanding the human myocardial small heat shock protein response may have significant implications for surgical myocardial protection.
Clements
Richard T
RT
Department of Surgery, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Mass, USA.
Sodha
Neel R
NR
Feng
Jun
J
Mieno
Shigetoshi
S
Boodhwani
Munir
M
Ramlawi
Basel
B
Bianchi
Cesario
C
Sellke
Frank W
FW
D008841:Actin Cytoskeleton / Q000378:metabolism / Q000473:pathology; D002315:Cardiopulmonary Bypass / Q000009:adverse effects*; D004591:Electrophoresis, Polyacrylamide Gel; D006324:Heart Arrest, Induced / Q000009:adverse effects*; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D015428:Myocardial Reperfusion Injury / Q000209:etiology / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D010766:Phosphorylation; D038203:alpha-Crystallin B Chain / Q000378:metabolism*
D006360:Heat-Shock Proteins; D038203:alpha-Crystallin B Chain