Switching ubiquitous and muscle‐specific isoforms of mitochondrial respiratory complex IV in skeletal muscle fine‐tunes complex IV activity

Abstract Respiratory complex IV (CIV, cytochrome c oxidase) is the terminal enzyme of the mitochondrial electron transport chain. Some CIV subunits have two or more isoforms, which are ubiquitously expressed or are expressed in specific tissues like the lung, muscle, and testis. Among the tissue‐specific CIV isoforms, the muscle‐specific isoforms are expressed in adult cardiac and skeletal muscles. To date, the physiological and biochemical association between the muscle‐specific CIV isoforms and aerobic respiration in muscles remains unclear. In this study, we profiled the CIV organization and expression pattern of muscle‐specific CIV isoforms in different mouse muscle tissues. We found extensive CIV‐containing supramolecular organization in murine musculature at advanced developmental stages, while a switch in the expression from ubiquitous to muscle‐specific isoforms of CIV was also detected. Such a switch was confirmed during the in vitro differentiation of mouse C2C12 myoblasts. Unexpectedly, a CIV expression decrease was observed during C2C12 differentiation, which was probably due to a small increase in the expression of muscle‐specific isoforms coupled with a dramatic decrease in the ubiquitous isoforms. We also found that the enzymatic activity of CIV containing the muscle‐specific isoform COX6A2 was higher than that with COX6A1 in engineered HEK293T cells. Overall, our results indicate that switching the expression from ubiquitous to muscle‐specific CIV isoforms is indispensable for optimized oxidative phosphorylation in mature skeletal muscles. We also note that the in vitro C2C12 differentiation model is not suitable for the study of muscular aerobic respiration due to insufficient expression of muscle‐specific CIV isoforms.