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PMID: 180974 Published · ppublish English Journal Article

Regulation of pyruvate dehydrogenase in rat heart. Mechanism of regulation of proportions of dephosphorylated and phosphorylated enzyme by oxidation of fatty acids and ketone bodies and of effects of diabetes: role of coenzyme A, acetyl-coenzyme A and reduced and oxidized nicotinamide-adenine dinucleotide.

The Biochemical journal ·Vol. 154 ·No. 2 ·1976-02-15 ·Pages 327-48

Kerbey AL, Randle PJ, Cooper RH, Whitehouse S, Pask HT, Denton RM

Abstract

The proportion of active (dephosphorylated) pyruvate dehydrogenase in perfused rat heart was decreased by alloxan-diabetes or by perfusion with media containing acetate, n-octanoate or palmitate. The total activity of the dehydrogenase was unchanged. 2. Pyruvate (5 or 25mM) or dichloroacetate (1mM) increased the proportion of active (dephosphorylated) pyruvate dehydrogenase in perfused rat heart, presumably by inhibiting the pyruvate dehydrogenase kinase reaction. Alloxan-diabetes markedly decreased the proportion of active dehydrogenase in hearts perfused with pyruvate or dichloroacetate. 3. The total activity of pyruvate dehydrogenase in mitochondria prepared from rat heart was unchanged by diabetes. Incubation of mitochondria with 2-oxo-glutarate plus malate increased ATP and NADH concentrations and decreased the proportion of active pyruvate dehydrogenase. The decrease in active dehydrogenase was somewhat greater in mitochondria prepared from hearts of diabetic rats than in those from hearts of non-diabetic rats. Pyruvate (0.1-10 mM) or dichloroacetate (4-50 muM) increased the proportion of active dehydrogenase in isolated mitochondria presumably by inhibition of the pyruvate dehydrogenase kinase reaction. They were much less effective in mitochondria from the hearts of diabetic rats than in those of non-diabetic rats. 4. The matrix water space was increased in preparations of mitochondria from hearts of diabetic rats. Dichloroacetate was concentrated in the matrix water of mitochondria of non-diabetic rats (approx. 16-fold at 10 muM); mitochondria from hearts of diabetic rats concentrated dichloroacetate less effectively. 5. The pyruvate dehydrogenase phosphate phosphatase activity of rat hearts and of rat heart mitochondria (approx. 1-2 munit/unit of pyruvate dehydrogenase) was not affected by diabetes. 6. The rate of oxidation of [1-14C]pyruvate by rat heart mitochondria (6.85 nmol/min per mg of protein with 50 muM-pyruvate) was approx. 46% of the Vmax. value of extracted pyruvate dehydrogenase (active form). Palmitoyl-L-carnitine, which increased the ratio of [acetyl-CoA]/[CoA] 16-fold, inhibited oxidation of pyruvate by about 90% without changing the proportion of active pyruvate dehydrogenase.

MeSH Terms
Acetates/pharmacology Acetyl Coenzyme A/metabolism Adenosine Triphosphate/metabolism Animals Caprylates/pharmacology Carnitine/analogs & derivatives,pharmacology Cell Membrane/metabolism Coenzyme A/metabolism Diabetes Mellitus, Experimental/metabolism Fatty Acids/metabolism Heart/drug effects In Vitro Techniques Ketoglutaric Acids/pharmacology Ketone Bodies/metabolism Malates/metabolism Male Mitochondria, Muscle/enzymology Myocardium/enzymology NAD/metabolism Oligomycins/metabolism Palmitates/pharmacology Perfusion Phosphates/metabolism Pyruvate Dehydrogenase Complex/metabolism Pyruvates/metabolism,pharmacology Rats Succinates/metabolism
Chemicals
Acetates Caprylates Fatty Acids Ketoglutaric Acids Ketone Bodies Malates Oligomycins Palmitates Phosphates Pyruvate Dehydrogenase Complex Pyruvates Succinates NAD Acetyl Coenzyme A Adenosine Triphosphate Carnitine Coenzyme A
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kerbey A L
Randle P J
Cooper R H
Whitehouse S
Pask H T
Denton R M
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj1540327
Published
1976-02-15
Pages
327-48
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1172714
Subset
IM
External Links
PubMed source DOI: 10.1042/bj1540327 PMC Full Text View journal details
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