Isolation and characterization of 3-hydroxyacyl coenzyme A dehydrogenase-binding protein from pig heart inner mitochondrial membrane

3-Hydroxyacyl coenzyme A (CoA) dehydrogenase-binding protein was solubilized from inner mitochondrial membrane by using taurodeoxycholate at high ionic strength. The binding protein was isolated from the suspension using 3-hydroxyacyl-CoA dehydrogenase affinity chromatography. The protein eluted fro...

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Bibliographic Details
Published in:The Journal of biological chemistry 1986-10, Vol.261 (30), p.14209-14213
Main Authors: Kispal, G, Sumegi, B, Alkonyi, I
Format: Article
Language:English
Subjects:
3-Hydroxyacyl CoA Dehydrogenases - metabolism
Analytical, structural and metabolic biochemistry
Animals
Biological and medical sciences
Carrier Proteins - isolation & purification
Enzymes and enzyme inhibitors
Fundamental and applied biological sciences. Psychology
Intracellular Membranes - analysis
Kinetics
Mitochondria, Heart - ultrastructure
Molecular Weight
Osmolar Concentration
Oxidoreductases
Swine
Taurodeoxycholic Acid - pharmacology
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Summary:3-Hydroxyacyl coenzyme A (CoA) dehydrogenase-binding protein was solubilized from inner mitochondrial membrane by using taurodeoxycholate at high ionic strength. The binding protein was isolated from the suspension using 3-hydroxyacyl-CoA dehydrogenase affinity chromatography. The protein eluted from the affinity column had a molecular weight of approximately 150,000, as determined by gel filtration. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the protein is a dimer consisting of 69,000 and 71,000 molecular weight subunits. The enzyme binding capacity of this protein was tested with a polyethylene glycol precipitation method: 0.5 mg of enzyme could be precipitated together with 1 mg of binding protein, showing that 1 mol of binding protein binds 1 mol of enzyme. This protein had no affinity toward malic dehydrogenase, citrate synthase, and fumarase. The approximately 2-fold increase in the 3-hydroxyacyl-CoA dehydrogenase activity when it was measured in the presence of the binding protein is additional evidence of enzyme-binding protein interaction. When incorporated into liposomes, the binding protein retained its ability to bind 3-hydroxyacyl-CoA dehydrogenase, but did not bind malic dehydrogenase, citrate synthase, and fumarase. These results suggest that the protein isolated by us has a specific function in anchoring a beta-oxidation enzyme to the matrix surface of the mitochondrial membrane.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(18)67005-X