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1.
Int J Mol Sci ; 21(23)2020 Nov 24.
Article in English | MEDLINE | ID: mdl-33255464

ABSTRACT

Small biomolecules, such as coenzyme A (CoA) and acetyl coenzyme A (acetyl-CoA), play vital roles in the regulation of cellular energy metabolism. In this paper, we evaluated the delayed effect of the potent hepatotoxin thioacetamide (TAA) on the concentrations of CoA and acetyl-CoA in plasma and in different rat tissues. Administration of TAA negatively affects liver function and leads to the development of hepatic encephalopathy (HE). In our experiments, rats were administered a single intraperitoneal injection of TAA at doses of 200, 400, or 600 mg/kg. Plasma, liver, kidney, and brain samples were collected six days after the TAA administration, a period that has been suggested to allow for restoration of liver function. The concentrations of CoA and acetyl-CoA in the group of rats exposed to different doses of TAA were compared to those observed in healthy rats. The results obtained indicate that even a single administration of TAA to rats is sufficient to alter the physiological balance of CoA and acetyl-CoA in the plasma and tissues of rats for an extended period of time. The initial concentrations of CoA and acetyl-CoA were not restored even after the completion of the liver regeneration process.


Subject(s)
Acetyl Coenzyme A/blood , Coenzyme A/blood , Hepatic Encephalopathy/blood , Thioacetamide/pharmacology , Acetyl Coenzyme A/genetics , Animals , Brain/drug effects , Brain/metabolism , Coenzyme A/genetics , Hepatic Encephalopathy/chemically induced , Hepatic Encephalopathy/pathology , Humans , Injections, Intraperitoneal , Liver/drug effects , Liver/pathology , Liver Regeneration/genetics , Rats , Thioacetamide/toxicity
2.
Nat Chem Biol ; 11(10): 784-92, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26322826

ABSTRACT

The metabolic cofactor coenzyme A (CoA) gained renewed attention because of its roles in neurodegeneration, protein acetylation, autophagy and signal transduction. The long-standing dogma is that eukaryotic cells obtain CoA exclusively via the uptake of extracellular precursors, especially vitamin B5, which is intracellularly converted through five conserved enzymatic reactions into CoA. This study demonstrates an alternative mechanism that allows cells and organisms to adjust intracellular CoA levels by using exogenous CoA. Here CoA was hydrolyzed extracellularly by ectonucleotide pyrophosphatases to 4'-phosphopantetheine, a biologically stable molecule able to translocate through membranes via passive diffusion. Inside the cell, 4'-phosphopantetheine was enzymatically converted back to CoA by the bifunctional enzyme CoA synthase. Phenotypes induced by intracellular CoA deprivation were reversed when exogenous CoA was provided. Our findings answer long-standing questions in fundamental cell biology and have major implications for the understanding of CoA-related diseases and therapies.


Subject(s)
Caenorhabditis elegans/metabolism , Coenzyme A/biosynthesis , Drosophila/metabolism , Pantetheine/analogs & derivatives , Animals , Caenorhabditis elegans/growth & development , Cell Line , Coenzyme A/blood , Coenzyme A/pharmacology , Coenzyme A Ligases/metabolism , Drosophila/cytology , Drosophila/growth & development , Female , HEK293 Cells , Humans , Longevity/physiology , Male , Mice, Inbred C57BL , Pantetheine/blood , Pantetheine/metabolism , Pantetheine/pharmacology , Phosphotransferases (Alcohol Group Acceptor)/genetics , Phosphotransferases (Alcohol Group Acceptor)/metabolism
3.
Molecules ; 22(9)2017 Aug 23.
Article in English | MEDLINE | ID: mdl-28832533

ABSTRACT

Coenzyme A (CoA) and acetyl-coenzyme A (acetyl-CoA) play essential roles in cell energy metabolism. Dysregulation of the biosynthesis and functioning of both compounds may contribute to various pathological conditions. We describe here a simple and sensitive HPLC-UV based method for simultaneous determination of CoA and acetyl-CoA in a variety of biological samples, including cells in culture, mouse cortex, and rat plasma, liver, kidney, and brain tissues. The limits of detection for CoA and acetyl-CoA are >10-fold lower than those obtained by previously described HPLC procedures, with coefficients of variation <1% for standard solutions, and 1-3% for deproteinized biological samples. Recovery is 95-97% for liver extracts spiked with Co-A and acetyl-CoA. Many factors may influence the tissue concentrations of CoA and acetyl-CoA (e.g., age, fed, or fasted state). Nevertheless, the values obtained by the present HPLC method for the concentration of CoA and acetyl-CoA in selected rodent tissues are in reasonable agreement with literature values. The concentrations of CoA and acetyl-CoA were found to be very low in rat plasma, but easily measurable by the present HPLC method. The method should be useful for studying cellular energy metabolism under normal and pathological conditions, and during targeted drug therapy treatment.


Subject(s)
Acetyl Coenzyme A/blood , Acetyl Coenzyme A/chemistry , Chromatography, High Pressure Liquid , Coenzyme A/blood , Coenzyme A/chemistry , Spectrophotometry, Ultraviolet , Animals , Cell Line , Cerebral Cortex/enzymology , Female , Humans , Mice , Rats
4.
Anal Sci ; 23(5): 557-61, 2007 May.
Article in English | MEDLINE | ID: mdl-17495401

ABSTRACT

A new spectrofluorometric method has been developed for the determination of trace amounts of coenzyme A (CoA). Using europium (Eu3+)-tetracycline (TC) complex as a fluorescent probe in the buffer solution of pH 6.80, CoA could remarkably enhance the fluorescence intensity of the Eu3+-TC complex at lambda = 612 nm after adding H(5)IO(6) and the enhanced fluorescence intensity is in proportion to the concentration of CoA. Optimum conditions for the determination of CoA were also investigated. The dynamic range for the determination of CoA is 6.08 x 10(-8) - 1.84 x 10(-5) mol L(-1) with detection limit of 4.62 x 10(-8) mol L(-1). This method is simple, practical and relatively free of interference from coexisting substances and can be successfully applied to determination of CoA in injection, human serum and pig liver samples. Moreover, the enhancement mechanisms of the fluorescence intensity in the Eu3+-TC system and the CoA-Eu3+-TC system have been also discussed.


Subject(s)
Coenzyme A/analysis , Coenzyme A/blood , Liver/chemistry , Spectrometry, Fluorescence/instrumentation , Swine , Animals , Coenzyme A/chemistry , Europium/chemistry , Humans , Hydrogen-Ion Concentration , Spectrometry, Fluorescence/methods , Spectrophotometry, Atomic , Tetracycline/chemistry
5.
Am J Clin Nutr ; 50(5): 1072-8, 1989 Nov.
Article in English | MEDLINE | ID: mdl-2816792

ABSTRACT

Reported normal concentrations for human whole-blood total pantothenic acid vary from 1.1 to 12 mumol/L. This wide range may partly arise from the various enzymes used for liberation of pantothenic acid from coenzyme A, particularly the source of pantetheinase. A purified pantetheinase from pig kidney had greater than 100 times the specific activity and less than 0.01 times the pantothenate content of other commonly used extracts. Endogenous pantetheinase activity in human plasma was identified (11.2 +/- 2.0 mumol pantothenate .min-1.L-1, n = 29) and found comparable to the activity usually added from exogenous sources for liberation of pantothenate from whole blood (1-13 mumol.min-1.L-1). Alkaline phosphatase alone liberated as much pantothenate from hemolyzed whole blood as did alkaline phosphatase with pantetheinase. Previous reports of total blood pantothenate may be elevated by pantothenate in the pantetheinase extracts, an unnecessary source of error. Whole-blood total pantothenate concentrations less than 4.6 mumol/L are normal and do not indicate deficiency, as is often currently quoted.


Subject(s)
Alkaline Phosphatase , Amidohydrolases , Coenzyme A/blood , Pantothenic Acid/blood , Animals , Chickens , Columbidae , GPI-Linked Proteins , Humans , Hydrolysis , Kidney/enzymology , Species Specificity , Swine , Temperature
6.
Am J Clin Nutr ; 67(3): 386-90, 1998 Mar.
Article in English | MEDLINE | ID: mdl-9497180

ABSTRACT

We studied the effects of L-carnitine treatment in the acyl flux of erythrocyte membranes from uremic patients. We found a significantly lower relative proportion of long-chain acyl-CoA (LCCoA) to free CoA (FCoA) in patients than in control subjects. In addition, patients had reduced activities of both carnitine palmitoyltransferase (CPT) and glycerophospholipid acyltransferase (LAT; CoA dependent), and increased ratios of long-chain acylcarnitine (LCAC) to free carnitine in their erythrocytes. These data support the hypothesis that acyl-trafficking is altered in erythrocytes in uremia. After treatment with L-carnitine, we observed a significant increase in CPT and LAT activities as well as in the LCCoA-FCoA ratio, and a significant decrease in the ratio of LCAC to free carnitine. These results support the conclusion that L-carnitine supplementation improves erythrocyte flux in uremic patients.


Subject(s)
Acyl Coenzyme A/blood , Carnitine/pharmacology , Coenzyme A/blood , Erythrocytes/drug effects , Sucrase/blood , Uremia/metabolism , Adult , Aged , Carnitine O-Palmitoyltransferase/metabolism , Electron Transport Complex IV/blood , Erythrocytes/enzymology , Erythrocytes/metabolism , Female , Humans , Male , Middle Aged , Uremia/enzymology
7.
Br J Pharmacol ; 122(3): 487-92, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9351505

ABSTRACT

1. Ibuprofen enantiomers and their respective coenzyme A thioesters were tested in human platelets and blood monocytes to determine their selectivity and potency as inhibitors of cyclo-oxygenase activity of prostaglandin endoperoxide synthase-1 (PGHS-1) and PGHS-2. 2. Human blood from volunteers was drawn and allowed to clot at 37 degrees C for 1 h in the presence of increasing concentrations of the test compounds (R-ibuprofen, S-ibuprofen, R-ibuprofenoyl-CoA, S-ibuprofenoyl-CoA, NS-398). Immunoreactive (ir) thromboxane B2 (TXB2) concentrations in serum were determined by a specific EIA assay as an index of the cyclo-oxygenase activity of platelet PGHS-1. 3. Heparin-treated blood from the same donors was incubated at 37 degrees C for 24 h with the same concentrations of the test compounds in the presence of lipopolysaccharide (LPS, 10 microg ml[-1]). The contribution of PGHS-1 was suppressed by pretreatment of the volunteers with aspirin (500 mg; 48 h before venepuncture). As a measure of LPS induced PGHS-2 activity immunoreactive prostaglandin E2 (irPGE2) plasma concentrations were determined by a specific EIA assay. 4. S-ibuprofen inhibited the activity of PGHS-1 (IC50 2.1 microM) and PGHS-2 (IC50 1.6 microM) equally. R-ibuprofen inhibited PGHS-1 (IC50 34.9) less potently than S-ibuprofen and showed no inhibition of PGHS-2 up to 250 microM. By contrast R-ibuprofenoyl-CoA thioester inhibited PGE2 production from LPS-stimulated monocytes almost two orders of magnitude more potently than the generation of TXB2 (IC50 5.6 vs 219 microM). 5. Western blotting of PGHS-2 after LPS induction of blood monocytes showed a concentration-dependent inhibition of PGHS-2 protein expression by ibuprofenoyl-CoA thioesters. 6. These data confirm that S-ibuprofen represents the active entity in the racemate with respect to cyclo-oxygenase activity. More importantly the data suggest a contribution of the R-enantiomer to therapeutic effects not only by chiral inversion to S-ibuprofen but also via inhibition of induction of PGHS-2 mediated by R-ibuprofenoyl-CoA thioester. 7. The data may explain why racemic ibuprofen is ranked as one of the safest non-steroidal anti-inflammatory drugs (NSAIDs) so far determined in epidemiological studies.


Subject(s)
Coenzyme A/pharmacology , Ibuprofen/pharmacology , Prostaglandin-Endoperoxide Synthases/blood , Blood Platelets/enzymology , Blotting, Western , Coenzyme A/blood , Cyclooxygenase 1 , Cyclooxygenase 2 , Cyclooxygenase 2 Inhibitors , Cyclooxygenase Inhibitors/pharmacology , Dinoprostone/blood , Esters , Humans , Ibuprofen/blood , In Vitro Techniques , Isoenzymes/blood , Membrane Proteins , Monocytes/enzymology , Stereoisomerism , Sulfides/blood , Sulfides/pharmacology , Thromboxane B2/blood
8.
Nutr Metab ; 18(3): 137-51, 1975.
Article in English | MEDLINE | ID: mdl-1187072

ABSTRACT

Plasma lecithin:cholesterol acyltransfer (LCAT), liver acyl-CoA:cholesterol acyltransfer (ACAT) and plasma and liver lipids were examined in newborn and young guinea pigs fed ordinary or a non-milk diet. LCAT in newborns was about 70% of the adult value, increased the first 3 days to values about three times the adult value, and decreased during the next 3 weeks to about adult value. Plasma lipids varied in a similar way. Liver ACAT in newborns was considerably higher than in adults. In young animals on non-milk diet the LCAT and ACAT were lower than in normally fed animals. LCAT was positively correlated to free cholesterol in plasma.


Subject(s)
Cholesterol/metabolism , Diet , Lipid Metabolism , Liver/metabolism , Milk , Triglycerides/metabolism , Acyltransferases/blood , Acyltransferases/metabolism , Animals , Cholesterol/blood , Cholesterol Esters/blood , Cholesterol Esters/metabolism , Coenzyme A/blood , Coenzyme A/metabolism , Esterification , Female , Glycerol/blood , Glycerol/metabolism , Guinea Pigs , Liver/enzymology , Male , Phosphatidylcholine-Sterol O-Acyltransferase/blood , Phosphatidylcholine-Sterol O-Acyltransferase/metabolism , Triglycerides/blood
9.
Clin Chim Acta ; 126(3): 307-13, 1982 Dec 23.
Article in English | MEDLINE | ID: mdl-7151284

ABSTRACT

The amounts of coenzyme A (CoASH) and acetyl-CoA in the acid-soluble extract of human blood platelets were quantitated by an isocratic reversed-phase liquid-chromatographic system. The analytical column was Supelcosil LC-18, and the solvent was 220 mmol/l potassium phosphate, pH 4.0, and 120 ml/l methanol. Long-chain acyl-CoA was estimated by the released coenzyme A after an alkaline hydrolysis of the acid-insoluble material of the blood platelets. The median values in platelets from fourteen healthy persons were 30 pmol CoASH/mg platelet protein, 45 pmol acetyl-CoA/mg platelet protein, and 34 pmol long-chain acyl-CoA/mg platelet protein.


Subject(s)
Acetyl Coenzyme A/blood , Acyl Coenzyme A/blood , Blood Platelets/analysis , Coenzyme A/blood , Carnitine O-Acetyltransferase , Chromatography, High Pressure Liquid , Humans , Phosphate Acetyltransferase
10.
Article in Russian | MEDLINE | ID: mdl-3577515

ABSTRACT

Sixty-three patients with the typical variant of delirium tremens were examined. The results showed a marked reduction (by 8.7 times) in levels of coenzyme A (CoA) in leukocytes, which was indicative of deficiency of the coenzymic form of pantothenic acid (PA). Changes in CoA concentrations in leukocytes observed at the height of psychosis in patients with alcoholic delirium as compared to other parameters of PA metabolism have great significance for the assessment of vitamin metabolism. PA deficiency was more expressed in cases of long-standing and massive alcoholization which induces an earlier development of psychosis. CoA levels in leukocytes may be used as a parameter of detoxication processes in patients with delirium tremens. The data obtained are considered as indication for administration of PA drugs in combined detoxifying therapy of alcoholic psychoses.


Subject(s)
Alcohol Withdrawal Delirium/metabolism , Coenzyme A/blood , Leukocytes/enzymology , Pantothenic Acid/analysis , Psychoses, Alcoholic/metabolism , Adult , Alcohol Withdrawal Delirium/enzymology , Alcohol Withdrawal Delirium/therapy , Alcoholism/blood , Combined Modality Therapy , Humans , Middle Aged , Pantothenic Acid/therapeutic use
11.
Vopr Onkol ; 45(5): 538-41, 1999.
Article in Russian | MEDLINE | ID: mdl-10629713

ABSTRACT

A relationship between blood antioxidant status in patients with cervical tumors (T3NxM0) and size of primary tumor has been investigated. Enzymatic and non-enzymatic components of antioxidant defenses were most severely damaged in cases of large tumors which suggested a specific suppression of adaptation systems by malignancies. These findings make a case for further research in antioxidant therapy for cervical carcinoma with due consideration of pathogenetic features of tumor disease.


Subject(s)
Antioxidants/metabolism , Uterine Cervical Neoplasms/blood , Uterine Cervical Neoplasms/pathology , Antioxidants/therapeutic use , Catalase/blood , Coenzyme A/blood , Female , Humans , Superoxide Dismutase/blood , Uterine Cervical Neoplasms/enzymology , Vitamin A/blood , Vitamin E/blood
18.
Nucleic Acids Res ; 9(19): 5061-73, 1981 Oct 10.
Article in English | MEDLINE | ID: mdl-7312625

ABSTRACT

We have studied histone acetylation in chicken erythrocytes. We find that about 30% of the histone in these cells is acetylated, however the majority of these histones are not in a dynamic steady state typical of other chicken cells and of mammalian cells, but rather are frozen in this state of modification. A very small fraction of erythrocyte histones are being modified normally but cannot be detected as shifting to higher levels of acetylation upon treatment with butyrate because the amount of histone so modified is small. Nonetheless, chicken erythrocytes incorporate 3H-acetate into histones about 40% as well as seen in the dynamically active HTC cells. This is most likely due to the formation of very high specific activity Acetyl CoA pools in erythrocytes which have very low levels of coenzyme A. We conclude that these genetically inactive cells are involved in only a minor way with histone acetylation.


Subject(s)
Butyrates/pharmacology , Erythrocytes/metabolism , Histones/blood , Acetates/blood , Acetyl Coenzyme A/blood , Acetylation , Animals , Butyric Acid , Chickens , Coenzyme A/blood , Rats
19.
J Biol Chem ; 259(4): 2403-6, 1984 Feb 25.
Article in English | MEDLINE | ID: mdl-6421811

ABSTRACT

Platelet membranes contain two distinct transacylase activities catalyzing the synthesis of arachidonoyl phosphatides by acylation of added lysophosphatides with endogenous esterified arachidonate. In the absence of CoA, arachidonate is incorporated only into ethanolamine lysophosphatides with a high preference for the plasmalogen form (Kramer, R. M., and Deykin, D. (1983) J. Biol. Chem. 258, 13806-13811). In the presence of CoA, however, lysophospholipids are acylated in the order 1-acyl-lysophosphatidylserine greater than 1-acyl-lysophosphatidylethanolamine greater than 1-acyl-lysophosphatidylinositol. The CoA-mediated transacylation reaction was characterized with 1-acyl-lysophosphatidylserine as acyl acceptor. It was highly specific for arachidonate and preferentially used phosphatidylcholine as the arachidonoyl donor. This enzymatic pathway may be part of a deacylation-transacylation cycle for remodeling of phospholipids synthesized de novo (according to the Lands pathway) representing a mechanism for enrichment of phospholipids with arachidonic acid.


Subject(s)
Arachidonic Acids/metabolism , Blood Platelets/metabolism , Coenzyme A/blood , Acylation , Arachidonic Acid , Cell Membrane/metabolism , Humans , Kinetics , Phospholipids/blood
20.
J Lipid Res ; 21(3): 292-300, 1980 Mar.
Article in English | MEDLINE | ID: mdl-7381324

ABSTRACT

Techniques have been developed to permit detection of acyl thioesters derived from exogenous fatty acids in erythrocytes. These acyl thioesters have been shown to act as intermediates in the acylation of endogenous lysophospholipid. Release of fatty acids from erythrocyte phospholipids has also been detected. Such release reflect the activity of an endogenous phospholipase that utilizes endogenous phospholipid as substrate. These observations permit further definition of the biochemical pathway for erythrocyte phospholipid fatty acid turnover.


Subject(s)
Erythrocyte Membrane/metabolism , Erythrocytes/metabolism , Fatty Acids/blood , Membrane Lipids/blood , Phospholipids/blood , Chemical Phenomena , Chemistry , Coenzyme A/analogs & derivatives , Coenzyme A/blood , Fatty Acids, Nonesterified/blood , Humans , Hydroxylamines , In Vitro Techniques
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