Calpain-1-sensitive myofibrillar proteins of the human myocardium.
Calpain-1 is a ubiquitous intracellular Ca2+-activated protease, which has been implicated in the pathogenesis of reversible myocardial depression (i.e. myocardial stunning) that follows ischemia and reperfusion via myofibrillar protein degradation. However, the target proteins of this degradative process in the human myocardium have not yet been identified. In order to compare the levels of Calpain-1 susceptibility within a set of human myofibrillar proteins (titin, alpha-fodrin, desmin, troponin T (cTnT), troponin I (cTnI) and alpha-actinin), crude left ventricular tissue homogenates were incubated for 0.5, 15, 30, 60 or 120 min in the presence of Calpain-1 (1 U or 5 U). Differences in the kinetics and extents of protein degradation were subsequently evaluated by using silver-stained SDS-polyacrylamide gels and Western immunoblot analyses. These assays revealed myofibrillar proteins with high (titin and alpha-fodrin), moderate (desmin and cTnT), or low (cTnI and alpha-actinin) relative Calpain-1 susceptibilities. The level of phosphorylation of cTnI did not explain its relatively low Calpain-1 susceptibility. Moreover, the molecular mass distributions of the truncated alpha-fodrin, desmin and cTnI fragments resulting from Ca2+-dependent autoproteolysis exhibited marked similarities with those of their Calpain-1-clipped products. These in vitro results shed light on a number of structural (titin, alpha-fodrin, desmin and alpha-actinin) and regulatory (cTnT and cTnI) proteins within the contractile apparatus as potential targets of Calpain-1. Their degradation may contribute to the development of postischemic stunning in the human myocardium.
Barta
Judit
J
Division of Clinical Physiology, Institute of Cardiology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary.
Tóth
Attila
A
Edes
István
I
Vaszily
Miklós
M
Papp
Julius Gy
JG
Varró
András
A
Papp
Zoltán
Z
D000185:Actinin / Q000378:metabolism; D000328:Adult; D000368:Aged; D002154:Calpain / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D064211:Connectin; D003893:Desmin / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D008840:Microfilament Proteins / Q000378:metabolism*; D008875:Middle Aged; D009124:Muscle Proteins / Q000378:metabolism; D009206:Myocardium / Q000201:enzymology / Q000378:metabolism*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D014336:Troponin / Q000378:metabolism; D013049:Spectrin
D000185:Actinin; D002154:Calpain; D002352:Carrier Proteins; D064211:Connectin; D003893:Desmin; D008840:Microfilament Proteins; D009124:Muscle Proteins; D011494:Protein Kinases; D014336:Troponin; D013049:Spectrin; C577285:TTN protein, human