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J. Biol. Chem., Vol. 265, Issue 36, 22204-22209, Dec, 1990
J Kopecky, M Baudysova, F Zanotti, D Janikova, S Pavelka and J Houstek
In order to characterize the biogenesis of unique thermogenic mitochondria
of brown adipose tissue, differentiation of precursor cells isolated from
mouse brown adipose tissue was studied in cell culture. Synthesis of
mitochondrial uncoupling protein (UCP), F1- ATPase, and cytochrome oxidase
was examined by L-[35S]methionine labeling and immunoblotting. For the
first time, synthesis of physiological amounts of the UCP, a key and
tissue-specific component of thermogenic mitochondria, was observed in
cultures at about confluence (day 6), indicating that a complete
differentiation of brown adipocytes was achieved in vitro. In postconfluent
cells (day 8) the content of UCP decreased rapidly, in contrast to some
other mitochondrial proteins (beta subunit of F1-ATPase, cytochrome
oxidase). In these cells, it was possible, by using norepinephrine, to
induce specifically the synthesis of the UCP but not of F1-ATPase or
cytochrome oxidase. The maximal response was observed at 0.1 microM
norepinephrine and the synthesis of UCP remained activated for at least 24
h. Detailed analysis revealed a major role of the beta-adrenergic receptors
and elevated intracellular concentration of cAMP in stimulation of UCP
synthesis. A quantitative recovery of the newly synthesized UCP in the
mitochondrial fraction indicated completed biogenesis of functionally
competent thermogenic mitochondria.
Synthesis of mitochondrial uncoupling protein in brown adipocytes differentiated in cell culture
Institute of Physiology, Czechoslovak Academy of Sciences, Prague.
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