Open AccessA monoclonal antibody to the Ca 2+ ‐ATPase of cardiac sarcoplasmic reticulum cross‐reacts with slow type I but not with fast type II canine skeletal muscle fibers: An immunocytochemical and immunochemical study
Author(s) -
Jorgensen Annelise O.,
Arnold Wayne,
Pepper David R.,
Kahl Steven D.,
Mandel Frederick,
Campbell Kevin P.
Publication year - 1988
Publication title -
cell motility and the cytoskeleton
Language(s) - English
Resource type - Journals
eISSN - 1097-0169
pISSN - 0886-1544
DOI - 10.1002/cm.970090208
Subject(s) - endoplasmic reticulum , skeletal muscle , biology , monoclonal antibody , cardiac muscle , microbiology and biotechnology , atpase , calcium atpase , immunocytochemistry , epitope , biochemistry , antibody , endocrinology , enzyme , immunology
Ca 2+ ‐ATPase of the sarcoplasmic reticulum was localized in cryostat sections from three different adult canine skeletal muscles (gracilis, extensor carpi radialis, and superficial digitalis flexor) by immunofluorescence labeling with monoclonal antibodies to the Ca 2+ ‐ATPase Type I (slow) myofibers were strongly labeled for the Ca 2+ ‐ATPase with a monoclonal antibody (II D8) to the CA 2+ ‐ATPase of canine cardiac sarcoplasmic reticulum; the type II (fast) myofibers were labeled at the level of the background with monoclonal antibody II D8. By contrast, type II (fast) myofibers were strongly labeled for Ca 2+ ‐ATPase of rabbit skeletal sarcoplasmic reticulum. The subcellular distribution of the immunolabeling in type I (slow) myofibers with monoclonal antibody II D8 corresponded to that of the sarcoplasmic reticulum as previously determined by electron microscopy. The structural similarity between the canine cardiac Ca 2+ ‐ATPase present in the sarcoplasmic reticulum of the canine slow skeletal muscle fibers was demonstrated by immunoblotting. Monoclonal antibody (II D8) to the cardiac Ca 2+ ‐ATPase binds to only one protein band present in the extract from either cardiac or type I (slow) skeletal muscle tissue. By contrast, monoclonal antibody (II H11) to the skeletal type II (fast) Ca 2+ ‐ATPase binds only one protein band in the extract from type II (fast) skeletal muscle tissue. These immunopositive proteins coelectrophoresed with the Ca 2+ ‐ATPase of the canine cardiac sarcoplasmic reticulum and showed an apparent M r of 115,000. It is concluded that the Ca 2+ ‐ATPase of cardiac and type I (slow) skeletal sarcoplasmic reticulum have at least one epitope in common, which is not present on the Ca 2+ ‐ATPase of sarcoplasmic reticulum in type II (fast) skeletal myofibers. It is possible that this site is related to the assumed necessity of the Ca 2+ ‐ATPase of the sarcoplasmic reticulum in cardiac and type I (slow) skeletal myofibers to interact with phosphorylated phospholamban and thereby enhance the accumulation of Ca 2+ in the lumen of the sarcoplasmic reticulum following β–adrenergic stimulation.