The calcium-release channel from cardiac sarcoplasmic reticulum: function in the failing and acutely ischaemic heart.
Junctional SR membrane vesicles have been isolated from chronically failing human hearts explanted at transplant operations. Vesicles have been incorporated into artificial planar phospholipid bilayers and the activity of single calcium-release channels investigated under voltage-clamp conditions. The properties of these channels are similar to those previously reported from normal animal tissue and do not provide evidence that the function of individual calcium-release channels is altered in the failing heart. Using radio-labelled ryanodine binding as a specific marker for the calcium-release channel, we demonstrate that, in the sheep heart, ischaemia results in the degradation of the calcium-release channel. The activation of proteases and oxidant stress in the ischaemic and re-perfused post-ischaemic myocardium are likely mediators of cell injury. Using the protease trypsin and the photosensitisation of rose bengal to generate the reactive oxygen species (ROS) singlet oxygen and superoxide radicals we demonstrate a direct effect on the calcium-release channel in vitro. Exposure of junctional SR vesicles to trypsin or oxidant stress resulted in the progressive loss of specific ryanodine binding and the degradation of high molecular weight proteins identified by polyacrylamide gel electrophoresis. The activity of single channels was also modified during exposure to proteolysis or oxidant stress; an initial increase in channel opening was observed followed by irreversible loss of channel function. Degradation of specific proteins, such as the calcium-release channel, may contribute to contractile dysfunction in the ischaemic and reperfused post-ischaemic myocardium.
Holmberg
S R
SR
Department of Cardiology, Royal Sussex County Hospital, Brighton, East Sussex, England.
Williams
A J
AJ
D000328:Adult; D000368:Aged; D000818:Animals; D015220:Calcium Channels / Q000187:drug effects / Q000378:metabolism*; D003327:Coronary Disease / Q000503:physiopathology*; D004195:Disease Models, Animal; D010450:Endopeptidases / Q000378:metabolism; D004789:Enzyme Activation; D006333:Heart Failure / Q000503:physiopathology; D006801:Humans; D008875:Middle Aged; D015427:Reperfusion Injury / Q000503:physiopathology; D012433:Ryanodine; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D012756:Sheep; D013481:Superoxides / Q000494:pharmacology*
D015220:Calcium Channels; D013481:Superoxides; D012433:Ryanodine; D010450:Endopeptidases