Calcium chloride in vitro effects on isolated myofibrillar proteins

The objective of this study was to determine the effect of 30 mM CaCl(2) on the solubilization of those structural proteins that contribute to myofibril stability. Ovine M. longissimus dorsi (longissimus) samples were obtained immediately post-exsanguination, myofibrils were isolated, glycerated, and frozen until needed. Myofibrils were washed, diluted and incubated in 0·1 m KCl, 10 mm Tris, pH 7·0 buffer for 24, 48 and 72 h. Treatments consisted of: (1) control, (2) 1 mm E(64), (3) 30 mm CaCl(2), and (4) 1 mm E(64) + 30 mm CaCl(2). Results (SDS-PAGE) indicated that myosin heavy chain (though not to a great extent), M-protein, C-protein, α-actinin, actin, troponin-T, tropomyosin isoforms, troponin-I and 72, 70, 62, 33, 32, 30, and 22 kDa unidentified bands were solubilized from myofibrils incubated in KCl buffer for 72 h. The addition of CaCl(2) hastened the appearance of some of the proteins in the supernatant fractions, but no differences were observed at 72 h among the treatments. The addition of E(64) had no effect on which proteins were released. Thus, in the absence of proteolysis it appears that a general solubilization of thick-and-thin filament ancillary proteins occurs in the presence of 30 mm CaCl(2). However, the contribution to tenderness should be minimal, because solubilized proteins are not part of the cytoskeletal elements that are responsible for maintaining structural integrity of the tissue.