Assay of short-chain acyl coenzyme a intermediates in tissue extracts by high-pressure liquid chromatography

Corkey, Barbara E.; Brandt, Martín; Williams, R. J.; Williamson, John

doi:10.1016/0003-2697(81)90152-4

Cited by 128 publications

(34 citation statements)

References 26 publications

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“…The propionyl-CoA peak was not seen when cell-free extract of strain JE4287 (vector-only control) was substituted for cell-free extract of strain JE4184 (pT7-6 prpE+) in the reaction mixture (data not shown). If the solvent system was changed to phosphate buffer with methanol (Corkey et al, 1981), both the product of the PrpE reaction and authentic propionyl-CoA eluted as a single peak (data not shown). This result suggested that multiple peaks observed in the acetonitrile/ammonium acetate solvent system were a mixture of different salts of the CoA moiety.…”

Section: Hplc Purification Of Prpe Productmentioning

confidence: 99%

The prpE gene of Salmonella typhimurium LT2 encodes propionyl-CoA synthetase

Horswill¹,

Escalante‐Semerena²

1999

Microbiology

109

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USABiochemical and genetic evidence is presented to demonstrate that the prpE gene of Salmonella typhimurium encodes propionyl-CoA synthetase, an enzyme required for the catabolism of propionate in this bacterium. While prpE mutants used propionate as carbon and energy source, prpE mutants that lacked acetyl-CoA synthetase (encoded by acs) did not, indicating that Acs can compensate for the lack of PrpE in prpE mutants. Cell-free extracts enriched for PrpE catalysed the formation of propionyl-CoA in a propionate-, ATP-, Mg2+-and HS-CoA dependent manner. Acetate substituted for propionate in the reaction at 48% the rate of propionate; butyrate was not a substrate for PrpE. The propionyl-CoA synthetase activity of PrpE was specific for ATP. GTP, ITP, CTP and TTP were not used as substrates by the enzyme. UV-visible spectrophotometry, HPLC and M S data demonstrated that propionyl-CoA was the product of the reaction catalysed by PrpE.

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Section: Hplc Purification Of Prpe Productmentioning

confidence: 99%

The prpE gene of Salmonella typhimurium LT2 encodes propionyl-CoA synthetase

Horswill¹,

Escalante‐Semerena²

1999

Microbiology

109

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“…Precipitated proteins were removed from the trichloroacetic acid extract by centrifugation and the supernatant was neutralized by extracting the acid five times with ether. Samples were lyophilized and stored at K70 8C until assayed (Corkey et al 1981(Corkey et al , 1988. Acid-soluble acyl-CoA compounds were determined by a modification of the reversedphase HPLC method of Corkey et al (1981Corkey et al ( , 1988.…”

Section: Determination Of Hypothalamic Malonyl-coamentioning

confidence: 99%

“…Samples were lyophilized and stored at K70 8C until assayed (Corkey et al 1981(Corkey et al , 1988. Acid-soluble acyl-CoA compounds were determined by a modification of the reversedphase HPLC method of Corkey et al (1981Corkey et al ( , 1988. This involved the use of acetonitrile rather than methanol as the organic component of the mobile phase, a flow rate of 0 .…”

Section: Determination Of Hypothalamic Malonyl-coamentioning

confidence: 99%

Intracerebroventricular injection of citrate inhibits hypothalamic AMPK and modulates feeding behavior and peripheral insulin signaling

Stoppa¹,

Cesquini²,

Roman³

et al. 2008

Journal of Endocrinology

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We hypothesized that citrate might modulate the AMP-activated protein kinase/acetyl-CoA carboxylase (AMPK)/(ACC) pathway and participate in neuronal feeding control and glucose homeostasis. To address this issue, we injected citrate into the lateral ventricle of rats. Intracerebroventricular (ICV) injection of citrate diminished the phosphorylation of hypothalamic AMPK/ACC, increased the expression of anorexigenic neuropeptide (pro-opiomelanocortin and corticotropinreleasing hormone), elevated the level of malonyl-CoA in the hypothalamus, and reduced food intake. No change was observed in the concentration of blood insulin after the injection of citrate. With a euglycemic-hyperinsulinemic clamp, the glucose infusion rate was higher in the citrate group than in the control group (28 . 6G0 . 8 vs 19 . 3G0 . 2 mU/kg body weight/min respectively), and so was glucose uptake in skeletal muscle and the epididymal fat pad. Concordantly, insulin receptor (IR), IR substrate type 1 (IRS1), IRS2, and protein kinase B (AKT) phosphorylation in adipose tissue and skeletal muscle was improved by citrate ICV treatment. Moreover, the treatment with citrate for 7 days promoted body weight loss and decreased the adipose tissue. Our results suggest that citrate and glucose may serve as signals of energy and nutrient availability to hypothalamic cells.

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“…Means in each row without a common superscript are significantly different at P<O pholipid enzymic kit (Boehringer Mannheim) and the Biopak Triacylglycerol enzymic kit (Biotrol). The VLDL triacylglycerol secretion rate was determined by measuring the increase in the plasma triacylglycerol concentration after an intravenous injection of Triton WR 1339 as described before [26, 271. Hepatic free CoASH and malonyl-CoA levels were determined by an HPLC method as described by Corkey et al [28].…”

Section: Chemicals and Drugsmentioning

confidence: 99%

Hepatic Fatty Acid Metabolism as a Determinant of Plasma and Liver Triacylglycerol Levels

Asiedu

Al‐Shurbaji

Rustan

et al. 1995

European Journal of Biochemistry

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To investigate the importance of factors influencing substrate availability for triacylglycerol biosynthesis on lipoprotein metabolism, the effects of two opposite-acting sulphur-substituted fatty acid analogues, tetradecylthioacetic acid and tetradecylthiopropionic acid were studied. Administration of tetradecylthioacetic acid to rats resulted in a reduction of plasma levels of triacylglycerols (44%) and cholesterol (26 %). This was accompanied by a reduction in very-low-density lipoprotein (VLDL) triacylglycerols (48 %), VLDL cholesterol (36% j, low-density lipoprotein (LDL) cholesterol (36 %) and high-density lipoprotein (HDL) triacylglycerols (50 %), whereas HDL cholesterol levels did not change. Subsequently, the HDLLDL-cholesterol ratio increased by 40 %. The cholesterol-lowering effect was accompanied by a reduction in hydroxymethylglutaryl CoA (HMG-CoA) reductase activity (37 %). Both mitochondrial and peroxisomal fatty acid oxidation increased (1.7-fold and 5.3-fold, respectively). Furthermore, there was a significant negative correlation between plasma triacylglycerols and mitochondrial fatty acid oxidation. Hepatic triacylglycerol synthesis was retarded, as indicated by a decrease in VLDL triacylglycerol secretion (40%), and by a reduced liver triacylglycerol content (29%). The activities of lipoprotein lipase and hepatic lipase in post-heparin plasma were not affected. Microsomal and cytosolic phosphatidate phosphohydrolase activities were inhibited (28 % and 70 %, respectively j. Hepatic malonyl-CoA levels decreased by 29% and the total activity of acetyl-CoA carboxylase was reduced (23%). In hepatocytes treated with tetradecylthioacetic acid, mitochondrial fatty acid oxidation increased markedly (100 %) and triacylglycerol secretion was reduced (40%). In tetradecylthiopropionic-acid-treated rats, a significant increase in both plasma and VLDL triacylglycerols was found (46 % and 72 %, respectively) but VLDL triacylglycerol secretion was unaffected. However, no effect on either plasma or lipoprotein cholesterol levels was seen. Mitochondrial fatty acid oxidation was decreased by 50 % and hepatic triacylglycerol levels increased by 33 %. In hepatocytes exposed to tetradecylthiopropionic acid, triacylglycerol synthesis increased (100 %) while triacylglycerol secretion and fatty acid oxidation remained unaltered. The results illustrate that lipoprotein triacylglycerol levels can be modulated by changes in the availability of fatty acid substrate for triacylglycerol biosynthesis, mainly by affecting mitochondrial fatty acid oxidation. In addition, we demonstrate that suppression of rat hepatic HMG-CoA reductase activity during treatment with tetradecylthioacetic acid may contribute to a cholesterol-lowering effect.

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Assay of short-chain acyl coenzyme a intermediates in tissue extracts by high-pressure liquid chromatography

Cited by 128 publications

References 26 publications

The prpE gene of Salmonella typhimurium LT2 encodes propionyl-CoA synthetase

The prpE gene of Salmonella typhimurium LT2 encodes propionyl-CoA synthetase

Intracerebroventricular injection of citrate inhibits hypothalamic AMPK and modulates feeding behavior and peripheral insulin signaling

Hepatic Fatty Acid Metabolism as a Determinant of Plasma and Liver Triacylglycerol Levels

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