Activities of enzymes involved in acetoacetate utilization in adult mammalian tissues

Abstract

1. The activities in rat tissues of 3-oxo acid CoA-transferase (the first enzyme involved in acetoacetate utilization) were found to be highest in kidney and heart. In submaxillary and adrenal glands the activities were about one-quarter of those in kidney and heart. In brain it was about one-tenth and was less in lung, spleen, skeletal muscle and epididymal fat. No activity was detectable in liver. 2. The activities of acetoacetyl-CoA thiolase were found roughly to parallel those of the transferase except for liver and adrenal glands. The high activity in the latter two tissues may be explained by additional roles of thiolase, namely, the production of acetyl-CoA from fatty acids. 3. The activities of the two enzymes in tissues of mouse, gerbil, golden hamster, guinea pig and sheep were similar to those of rat tissues. The notable exception was the low activity of the transferase and thiolase in sheep heart and brain. 4. The activities of the transferase in rat tissues did not change appreciably in starvation, alloxan-diabetes or on fat-feeding, where the rates of ketone-body utilization are increased. Thiolase activity increased in kidney and heart on fat-feeding. 5. The activity of 3-hydroxybutyrate dehydrogenase did not change in rat brain during starvation. 6. The factors controlling the rate of ketone-body utilization are discussed. It is concluded that the activities of the relevant enzymes in the adult rat do not control the variations in the rate of ketone-body utilization that occur in starvation or alloxan-diabetes. The controlling factor in these situations is the concentration of the ketone bodies in plasma and tissues.

Failure of systemic ketosis to control cachexia and the growth rate of the Walker 256 carcinosarcoma in rats

“…A similar distribution of enzyme activities has been reported for other tumours of the peripheral tissues (Tisdale & Brennan, 1983). Since activity of the enzyme 3-ketoacid CoA transferase determines the extent to which 3-hydroxybutyrate is used as a metabolic fuel (Williamson et al, 1971) it might be expected that the Walker 256 carcinosarcoma would be unable to metabolize ketone bodies. However, we have shown previously that Walker 256 tumour cells grown in vitro can form 14CO2 from D(-)3-hydroxy (3-14C-) butyrate, but only at a very low rate due to the presence of acetoacetyl CoA synthetase (Tisdale, 1984).…”

A simple purification of acetoacetate—succinate CoA‐transferase using substrate elution chromatography

“…Activity of CoA-transferase was measured spectrophotometrically in the direction of acetoacetyl-CoA breakdown by a modification of the method of Williamson et al [8]. The decrease in absorbance at 303 nm was followed in a 2 ml assay mixture at 30°C containing 100 mM Tris-sulphate (pH 8.1), 25 mM MgSO 4, 10 mM sodium succinate, 50/~M acetoacetylCoA and a suitable amount of the enzyme.…”

Fuel utilization in colonocytes of the rat

“…Assay of enzyme activities Enzyme activities were measured as described in the cited references: hexokinase ; glucose-6-phosphatase (Lackner et al, 1984); 6-phosphofructokinase (Opie & Newsholme, 1967); fructose-bisphosphatase 3-oxoacid CoA-transferase, acetoacetyl-CoA thiolase and D-3-hydroxybutyrate dehydrogenase (Williamson et al, 1971); glutaminase (Curthoys & Lowry, 1973); glutamate dehydrogenase ; alanine aminotransferase and aspartate aminotransferase (Sugden & Newsholme, 1975). The final volume of assay mixtures in all cases was 1.0 ml.…”