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Publication
Multiple mass isotopomer tracing of acetyl-CoA metabolism in
Langendorff-perfused rat hearts: channeling of acetyl-CoA from pyruvate
dehydrogenase to carnitine acetyltransferase.
Authors Li Q, Deng S, Ibarra RA, Anderson VE, Brunengraber H, Zhang GF
Submitted By Submitted Externally on 7/24/2015
Status Published
Journal The Journal of biological chemistry
Year 2015
Date Published 3/27/2015
Volume : Pages 290 : 8121 - 8132
PubMed Reference 25645937
Abstract We developed an isotopic technique to assess mitochondrial acetyl-CoA turnover
(˜citric acid flux) in perfused rat hearts. Hearts are perfused with buffer
containing tracer [(13)C2,(2)H3]acetate, which forms M5 + M4 + M3 acetyl-CoA.
The buffer may also contain one or two labeled substrates, which generate M2
acetyl-CoA (e.g. [(13)C6]glucose or [1,2-(13)C2]palmitate) or/and M1 acetyl-CoA
(e.g. [1-(13)C]octanoate). The total acetyl-CoA turnover and the contributions
of fuels to acetyl-CoA are calculated from the uptake of the acetate tracer and
the mass isotopomer distribution of acetyl-CoA. The method was applied to
measurements of acetyl-CoA turnover under different conditions (glucose ±
palmitate ± insulin ± dichloroacetate). The data revealed (i) substrate cycling
between glycogen and glucose-6-P and between glucose-6-P and triose phosphates,
(ii) the release of small excess acetyl groups as acetylcarnitine and ketone
bodies, and (iii) the channeling of mitochondrial acetyl-CoA from pyruvate
dehydrogenase to carnitine acetyltransferase. Because of this channeling, the
labeling of acetylcarnitine and ketone bodies released by the heart are not
proxies of the labeling of mitochondrial acetyl-CoA.




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