We propose to study the binding affinity of long chain fatty acyl CoA esters to and their effect on the adenine nucleotide translocase under in vivo conditions. For the experimental models we will use intact liver and heart, isolated perfused organs and isolated cells. Radioactive fatty acids included in the medium will be taken up by the tissues, metabolized to their acyl CoA esters, and binding of the acyl CoA to the translocase determined by isolating and purifying the acyl CoA-protein complex from mitochondrial extracts. The purification is essentially a one-step procedure using hydroxyapatite chromotography which permits the recovery of a homogeneous protein. The radioactive ligand is attached to the protein which thereby permits its analysis. The acyl CoA binding capacity will be compared in diabetic and normal livers and related to the adenine nucleotide translocase activity as well as the redox state and phosphate potential of the cell. These studies will provide evidence that cell metabolism is regulated at the level of adenine nucleotide translocation by long chain fatty acyl CoA esters.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM014033-18S1
Application #
3268589
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1979-01-01
Project End
1986-01-31
Budget Start
1985-09-23
Budget End
1986-01-31
Support Year
18
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
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Woldegiorgis, G; Duff, T; Contreras, L et al. (1989) Photoaffinity labeling of hamster brown adipose tissue mitochondria by an [125I] coenzyme A derivative: differential interaction with the uncoupling protein and ADP/ATP carrier. Biochem Biophys Res Commun 161:502-7
Molaparast-Saless, F; Shrago, E; Spennetta, T L et al. (1988) Determination of individual long-chain fatty acyl-CoA esters in heart and skeletal muscle. Lipids 23:490-2
Kim, J H; Woldgiorgis, G; Elson, C E et al. (1988) Age-related changes in respiration coupled to phosphorylation. I. Hepatic mitochondria. Mech Ageing Dev 46:263-77
Mares-Perlman, J A; Shrago, E (1988) Energy substrate utilization in freshly isolated Morris Hepatoma 7777 cells. Cancer Res 48:602-8

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