Mode of degradation of myofibrillar proteins by an endogenous protease, cathepsin L

Abstract

The mode of degradation of myofibrils and their constituent proteins by cathepsin L (EC 3.4.22.15) of rabbit skeletal muscle was studied. Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis showed that cathepsin L degraded myosin heavy chain, α-actinin, actin, troponin T and troponin I assembled in myofibrils and produced mainly fragments of 160 000 and 30 000 daltons in the acidic pH region. This degradation was most intense around pH 4. Degradation of myosin in the isolated state by cathepsin L resulted in the disappearance of the heavy chain and the decrease of light chains 1, 2 and 3, producing fragments of 160 000, 92 000, 83 000 and 60 000 daltons. The degradation of the heavy chain was most severe at pH 4.2. Cathepsin L degraded actin into fragments of 40 000, 37 000 and 30 000 daltons. This action was most intense at pH 4.7. Tropomyosin was not degraded. Troponin T and troponin I were degraded into fragments of 30 000 and 13 000 daltons at pH 3.7-6.7, which were degraded further into smaller fragments. Troponin C was not degraded. α-Actinin was degraded into several fragments, the major one of which showed an $\text{M}{}_{\mathrm{<mtext>r</mtext>}}$ of 80 000. This degradation was most intense at pH 3.0–3.5.