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Activation of pyruvate dehydrogenase by direct addition of insulin to an isolated plasma membrane/mitochondria mixture: evidence for generated of insulin's second messenger in a subcellular system.

Abstract：

:The addition of insulin to a mixture of plasma membrane and mitochondrial fractions from rat adipocytes results in a decrease in the phosphorylation of a mitochondrial protein identified as the alpha subunit of pyruvate dehydrogenase [pyruvate:lipoamide oxidoreductase (decarboxylating and acceptor-acetylating), EC 1.2.4.1] (Seals, J. R., McDonald, J. M. & Jarett, L. (1979) J. Biol. Chem. 254, 6991-6996). This study confirms the prediction that a corresponding increase in pyruvate dehydrogenase activity can be effected by insulin treatment of this preparation. Incubation of the plasma membrane/mitochondria mixture with ATP inhibited pyruvate dehydrogenase activity as measured in a subsequent enzyme assay. The presence of insulin during this incubation with ATP resulted in a 24.5% stimulation of enzyme activity compared to incubation without insulin (n = 9, P < 0.001). The effect was specific for biologically active insulin and was insulin dose-dependent in the physiological range of insulin. Supermaximal doses of insulin produced reduced effects. An insulin effect of similar magnitude could also be observed when the plasma membrane/mitochondria mixture was incubated without ATP. Two insulin mimickers, concanavalin A and antibody to insulin receptor, stimulated pyruvate dehydrogenase by 30.4% (n = 6, P <0.001) and 28.1% (n = 8, P<0.001), respectively. Both of these agents also produced reduced effects at supermaximal concentrations. The effects of all three agents required plasma membranes and could not be produced by treatment of mitochondria alone. The results suggest that a mechanism common to all three agents is responsible for transmitting the stimulation from the plasma membrane to the mitochondrial components of the mixture.

journal_name

Proc Natl Acad Sci U S A

journal_title

Proceedings of the National Academy of Sciences of the United States of America

authors

Seals JR,Jarett L

doi

10.1073/pnas.77.1.77

subject

Has Abstract

pub_date

1980-01-01 00:00:00

pages

77-81

issue

eissn

0027-8424

issn

1091-6490

journal_volume

pub_type

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Activation of pyruvate dehydrogenase by direct addition of insulin to an isolated plasma membrane/mitochondria mixture: evidence for generated of insulin's second messenger in a subcellular system.