Synthesis of coenzyme a esters of some bile acids

Shah, Parag P.; Staple, Ezra

doi:10.1016/s0039-128x(68)80034-0

Cited by 47 publications

(13 citation statements)

References 8 publications

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“…We detected the cholate-, CoA-, ATP-, and cell extract-dependent formation of a product, P5, which showed UV spectra similar to CoA. In addition, the UV spectrum of P5 showed a slight shoulder at 233 nm, which is characteristic for CoA esters (31). P5 had the same UV absorption spectrum and retention time in HPLC analysis as chemically synthesized cholyl-CoA (Fig.…”

Section: Lc-ms Analysis Revealed Deprotonated Molecular Ions ([M-h]mentioning

confidence: 78%

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Biochemical and Genetic Investigation of Initial Reactions in Aerobic Degradation of the Bile Acid Cholate in Pseudomonas sp. Strain Chol1

et al. 2007

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Bile acids are surface-active steroid compounds with toxic effects for bacteria. Recently, the isolation and characterization of a bacterium, Pseudomonas sp. strain Chol1, growing with bile acids as the carbon and energy source was reported. In this study, initial reactions of the aerobic degradation pathway for the bile acid cholate were investigated on the biochemical and genetic level in strain Chol1. These reactions comprised A-ring oxidation, activation with coenzyme A (CoA), and ␤-oxidation of the acyl side chain with the C 19 -steroid dihydroxyandrostadienedione as the end product. A-ring oxidizing enzyme activities leading to ⌬ 1,4 -3-ketocholyl-CoA were detected in cell extracts and confirmed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Cholate activation with CoA was demonstrated in cell extracts and confirmed with a chemically synthesized standard by LC-MS/MS. A transposon mutant with a block in oxidation of the acyl side chain accumulated a steroid compound in culture supernatants which was identified as 7␣,12␣-dihydroxy-3-oxopregna-1,4-diene-20-carboxylate (DHOPDC) by nuclear magnetic resonance spectroscopy. The interrupted gene was identified as encoding a putative acyl-CoA-dehydrogenase (ACAD). DHOPDC activation with CoA in cell extracts of strain Chol1 was detected by LC-MS/MS. The growth defect of the transposon mutant could be complemented by the wild-type ACAD gene located on the plasmid pBBR1MCS-5. Based on these results, the initiating reactions of the cholate degradation pathway leading from cholate to dihydroxyandrostadienedione could be reconstructed. In addition, the first bacterial gene encoding an enzyme for a specific reaction step in side chain degradation of steroid compounds was identified, and it showed a high degree of similarity to genes in other steroid-degrading bacteria.

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Section: Lc-ms Analysis Revealed Deprotonated Molecular Ions ([M-h]mentioning

confidence: 78%

“…Cholyl-CoA was synthesized via a mixed anhydride procedure as described previously (31). Formation of 14.2 mg of cholyl-CoA from 30 mg of sodium cholate (Fluka) and 50 mg of CoA (trilithium salt from yeast; Sigma) was confirmed by LC-MS/MS.…”

Section: Methodsmentioning

confidence: 99%

Biochemical and Genetic Investigation of Initial Reactions in Aerobic Degradation of the Bile Acid Cholate in Pseudomonas sp. Strain Chol1

et al. 2007

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“…Chemicals- [24-14 C]Chenodeoxycholoyl-CoA and C]choloylCoA (specific activity 48.6 Ci/mol), trihydroxy-cholestanoyl-CoA, 2-methylstearoyl-CoA, and prostaglandin F 2␣ -CoA were synthesized by the mixed anhydride procedure (29) and the former two CoA esters purified by high-pressure liquid chromatography (HPLC). [24-14 C]Chenodeoxycholic acid and C]cholic acid were purchased from PerkinElmer Life Sciences.…”

Section: Methodsmentioning

confidence: 99%

Characterization of an Acyl-CoA Thioesterase That Functions as a Major Regulator of Peroxisomal Lipid Metabolism

Hunt

Solaas

Kase

et al. 2002

Journal of Biological Chemistry

144

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Peroxisomes function in ␤-oxidation of very long and long-chain fatty acids, dicarboxylic fatty acids, bile acid intermediates, prostaglandins, leukotrienes, thromboxanes, pristanic acid, and xenobiotic carboxylic acids. These lipids are mainly chain-shortened for excretion as the carboxylic acids or transported to mitochondria for further metabolism. Several of these carboxylic acids are slowly oxidized and may therefore sequester coenzyme A (CoASH). To prevent CoASH sequestration and to facilitate excretion of chain-shortened carboxylic acids, acyl-CoA thioesterases, which catalyze the hydrolysis of acyl-CoAs to the free acid and CoASH, may play important roles. Here we have cloned and characterized a peroxisomal acyl-CoA thioesterase from mouse, named PTE-2 (peroxisomal acyl-CoA thioesterase 2). PTE-2 is ubiquitously expressed and induced at mRNA level by treatment with the peroxisome proliferator WY-14,643 and fasting. Induction seen by these treatments was dependent on the peroxisome proliferator-activated receptor ␣. Recombinant PTE-2 showed a broad chain length specificity with acyl-CoAs from short-and medium-, to long-chain acyl-CoAs, and other substrates including trihydroxycoprostanoyl-CoA, hydroxymethylglutaryl-CoA, and branched chain acyl-CoAs, all of which are present in peroxisomes. Highest activities were found with the CoA esters of primary bile acids choloyl-CoA and chenodeoxycholoyl-CoA as substrates. PTE-2 activity is inhibited by free CoASH, suggesting that intraperoxisomal free CoASH levels regulate the activity of this enzyme. The acyl-CoA specificity of recombinant PTE-2 closely resembles that of purified mouse liver peroxisomes, suggesting that PTE-2 is the major acyl-CoA thioesterase in peroxisomes. Addition of recombinant PTE-2 to incubations containing isolated mouse liver peroxisomes strongly inhibited bile acidCoA:amino acid N-acyltransferase activity, suggesting that this thioesterase can interfere with CoASH-dependent pathways. We propose that PTE-2 functions as a key regulator of peroxisomal lipid metabolism.Peroxisomes are cellular organelles present in all eukaryotic cells. They play an indispensable role in the metabolism of a variety of lipids including very long-chain fatty acids, dicarboxylic fatty acids, bile acids, prostaglandins, leukotrienes, thromboxanes, pristanic acid, and xenobiotic fatty acids (for review, see Refs. 1 and 2). The peroxisomal ␤-oxidation system contains two sets of enzymes, one of which is involved in the oxidation of branched chain fatty acids and intermediates in the hepatic bile acid biosynthetic pathway and consists of one or two branched-chain acyl-CoA oxidase(s), a D-specific bifunctional protein and the sterol carrier-like protein x (SCPx). The second pathway is involved in the oxidation of very long straight-chain fatty acids, CoA esters of prostaglandins, leukotrienes, and thromboxanes (prostanoids) and dicarboxylic acids. This system is composed of a straight-chain acyl-CoA oxidase, an L-specific bifunctional protein and straight-chain 3-ke...

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“…Cholyl-CoA was chemically synthesized as described previously (19), using a modification of the method of Shah and Staple (20). Cholyl-CoA was first separated from the reaction mixture by three extractions with five reaction volumes of ether in a separating funnel.…”

Section: Cholyl-coa Synthesismentioning

confidence: 99%

Conserved Residues in the Putative Catalytic Triad of Human Bile Acid Coenzyme A:Amino Acid N-Acyltransferase

Sfakianos¹,

Wilson

Sakalian

et al. 2002

Journal of Biological Chemistry

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Human bile acid-CoA:amino acid N-acyltransferase (hBAT), an enzyme catalyzing the conjugation of bile acids with the amino acids glycine or taurine has significant sequence homology with dienelactone hydrolases and other ␣/␤ hydrolases. These enzymes have a conserved catalytic triad that maps onto the mammalian BATs at residues Cys-235, Asp-328, and His-362 of the human sequence, albeit that the hydrolases contain a serine instead of a cysteine. In the present study, the function of the putative catalytic triad of hBAT was examined by chemical modification with the cysteine alkylating reagent N-ethylmaleimide (NEM) and by sitedirected mutagenesis of the triad residues followed by enzymology studies of mutant and wild-type hBATs.

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Synthesis of coenzyme a esters of some bile acids

Cited by 47 publications

References 8 publications

Biochemical and Genetic Investigation of Initial Reactions in Aerobic Degradation of the Bile Acid Cholate in Pseudomonas sp. Strain Chol1

Biochemical and Genetic Investigation of Initial Reactions in Aerobic Degradation of the Bile Acid Cholate in Pseudomonas sp. Strain Chol1

Characterization of an Acyl-CoA Thioesterase That Functions as a Major Regulator of Peroxisomal Lipid Metabolism

Conserved Residues in the Putative Catalytic Triad of Human Bile Acid Coenzyme A:Amino Acid N-Acyltransferase

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