Metabolic consequences of functional complexes of mitochondria, myofibrils and sarcoplasmic reticulum in muscle cells.
Regulation of mitochondrial respiration both by endogenous and exogenous ADP in the cells in situ was studied in isolated and permeabilized cardiomyocytes, permeabilized cardiac fibers and 'ghost' fibers (all with a diameter of 10-20 micro m) at different (0-3 micro moll(-1)) free Ca(2+) concentrations in the medium. In all these preparations, the apparent K(m) of mitochondrial respiration for exogenous ADP at free Ca(2+) concentrations of 0-0.1 micro moll(-1) was very high, in the range of 250-350 micro moll(-1), in contrast to isolated mitochondria in vitro (apparent K(m) for ADP is approximately 20 micro moll(-1)). An increase in the free Ca(2+) concentration (up to 3 micro moll(-1), which is within physiological range), resulted in a very significant decrease of the apparent K(m) value to 20-30 micro moll(-1), a decrease of V(max) of respiration in permeabilized intact fibers and a strong contraction of sarcomeres. In ghost cardiac fibers, from which myosin was extracted but mitochondria were intact, neither the high apparent K(m) for ADP (300-350 micro moll(-1)) nor V(max) of respiration changed in the range of free Ca(2+) concentration studied, and no sarcomere contraction was observed. The exogenous-ADP-trapping system (pyruvate kinase + phosphoenolpyruvate) inhibited endogenous-ADP-supported respiration in permeabilized cells by no more than 40%, and this inhibition was reversed by creatine due to activation of mitochondrial creatine kinase. These results are taken to show strong structural associations (functional complexes) among mitochondria, sarcomeres and sarcoplasmic reticulum. Inside these complexes, mitochondrial functional state is controlled by channeling of ADP, mostly via energy- and phosphoryl-transfer networks, and apparently depends on the state of sarcomere structures.
Andrienko
T
T
Laboratory of Fundamental and Applied Bioenergetics, INSERM E0221, Joseph Fourier University, Grenoble, France.
Kuznetsov
A V
AV
Kaambre
T
T
Usson
Y
Y
Orosco
A
A
Appaix
F
F
Tiivel
T
T
Sikk
P
P
Vendelin
M
M
Margreiter
R
R
Saks
V A
VA
D000244:Adenosine Diphosphate / Q000378:metabolism; D000818:Animals; D019069:Cell Respiration / Q000502:physiology; D007700:Kinetics; D008856:Microscopy, Fluorescence; D008929:Mitochondria, Heart / Q000378:metabolism*; D032342:Muscle Cells / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000378:metabolism; D009210:Myofibrils / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D012518:Sarcomeres / Q000378:metabolism; D012519:Sarcoplasmic Reticulum / Q000378:metabolism*
D000244:Adenosine Diphosphate