Striking Differences Between the Kinetics of Regulation of Respiration by ADP in Slow‐Twitch and Fast‐Twitch Muscles In Vivo

AV Kuznetsov, T Tiivel, P Sikk… - European Journal of …, 1996 - Wiley Online Library
AV Kuznetsov, T Tiivel, P Sikk, T Kaambre, L Kay, Z Daneshrad, A Rossi, L Kadaja, N Peet…
European Journal of Biochemistry, 1996Wiley Online Library
The kinetics of in vivo regulation of mitochondrial respiration by ADP was studied in rat
heart, slow‐twitch skeletal muscle (soleus) and fast‐twitch skeletal muscle (gastrocnemius,
plantaris, quadriceps and tibialis anterior) by means of saponin‐skinned fibres.
Mitochondrial respiratory parameters were determined in the absence and presence of
creatine (20 mM), and the effect of proteolytic enzymes (trypsin, chymotrypsin or elastase) on
these parameters was investigated in detail. The results of these experiments confirm the …
The kinetics of in vivo regulation of mitochondrial respiration by ADP was studied in rat heart, slow‐twitch skeletal muscle (soleus) and fast‐twitch skeletal muscle (gastrocnemius, plantaris, quadriceps and tibialis anterior) by means of saponin‐skinned fibres. Mitochondrial respiratory parameters were determined in the absence and presence of creatine (20 mM), and the effect of proteolytic enzymes (trypsin, chymotrypsin or elastase) on these parameters was investigated in detail. The results of these experiments confirm the observation of Veksler et al. [Veksler, V. I., Kuznetsov, A. V., Anflous, K., Mateo, P., van Deursen, J., Wieringa, B. & Ventura‐Clapier, R. (1995)J. Biol. Chem. 270, 19921–19929], who studied muscle fibres from normal and transgenic mice, that the kinetics of respiration regulation in muscle cells is tissue specific. We found that in rat cardiac and soleus muscle fibres the apparent Km for respiration regulation was 300–400 μM and decreased to 50–80 μM in the presence of creatine. In contrast, in skinned fibres from gastrocnemius, plantaris, tibialis anterior and quadriceps muscles, this value was initially very low, 10–20 μM, i.e. the same as that is in isolated muscle mitochondria, and the effect of creatine was not observable under these experimental conditions. Treatment of the fibres with trypsin, chymotrypsin or elastase (0.125 μg/ml) for 15 min decreased the apparent Km for ADP in cardiac and soleus muscle fibres to 40–98 μM without significant alteration of Vmax or the intactness of outer mitochondrial membrane, as assessed by the cytochrome c test. In fibres from gastrocnemius, trypsin increased the apparent Km, for ADP transiently. The effects of trypsin and chymotrypsin were studied in detail and found to be concentration dependent and time dependent. The effects were characterised by saturation phenomenon with respect to the proteolytic enzyme concentration, saturation being observed above 1 μM enzyme. These results are taken to show that in cardiac and slow‐twitch skeletal muscle, the permeability of the outer mitochondrial membrane to adenine nucleotides is low and controlled by a cytoplasmic protein that is sensitive to trypsin and chymotrypsin. This protein may participate in feedback signal transduction by a mechanism of vectorial‐ligand conduction. This protein factor is not expressed in fast‐twitch skeletal muscle, in which cellular mechanism of regulation of respiration is probably very different from that of slow‐twitch muscles.
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