Identification of a satellite fatty acid ethyl ester synthase from human myocardium as a glutathione S-transferase.

Bora, Puran S.; Spilburg, Curtis A.; Lange, Louis G.

doi:10.1172/jci114382

Cited by 36 publications

(22 citation statements)

References 20 publications

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“…Human myocardial FAEE synthase was purified to homogeneity by sequential DEAE-cellulose, gel permeation, hydrophobic interaction, and immunoaffinity chromatographies (1,6). In brief, human heart soluble fraction was applied to a DEAE-cellulose column (5 x 16 cm) in 1 mM 2-mercaptoethanol/10 mM Tris(hydroxymethyl)aminomethane (Tris), pH 8.0.…”

Section: Methodsmentioning

confidence: 99%

“…We have purified both a minor (cationic) and a major (anionic) form of FAEE synthase from human myocardium (6). The anionic form of FAEE synthase has a molecular weight of 26,000 and is the counterpart of that from rabbit heart (1).…”

mentioning

confidence: 99%

“…Enzyme Assays. Rates of FAEE synthesis were determined by the procedure described elsewhere (1,6). GSH transferase was assayed with 1-chloro-2,4-dinitrobenzene as substrate as described by Habig et al (12).…”

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confidence: 99%

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Metabolism of ethanol and carcinogens by glutathione transferases.

Bora

Spilburg²,

Lange³

1989

Proc. Natl. Acad. Sci. U.S.A.

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Nonoxidative alcohol metabolism to form fatty acid ethyl esters contributes to alcohol-related end-organ damage, and these products are formed by two synthase enzymes. We recently purified the major (pI 4.9) synthase from human myocardium. The N-terminal sequence is >73% identical with that of a neutral (pI 6.7) detoxification enzyme, glutathione transferase P from rat hepatoceliular carcinoma ( Previous reports characterized a metabolic pathway for ethanol in extrahepatic organs (1, 2) that we now relate to an important pathway for metabolism of carcinogens in liver. Brain, heart, and other organs lack oxidative ethanol metabolism and yet they are often injured by alcohol abuse. The observation that fatty acid ethyl esters (FAEEs) are synthesized at high rates in these tissues provided a plausible link between the observed tissue damage and the ingestion of alcohol (2). Moreover, these neutral lipids are potentially toxic for mitochondria (3) and, therefore, FAEEs may account in part for extrahepatic patterns of selective end-organ damage due to alcohol abuse. Recent genetic studies using human peripheral leukocytes have shown that this synthase activity is heritable in an autosomal recessive pattern for high activity (4,5).We have purified both a minor (cationic) and a major (anionic) form of FAEE synthase from human myocardium (6). The anionic form of FAEE synthase has a molecular weight of 26,000 and is the counterpart of that from rabbit heart (1). With these purified preparations, free fatty acid was found to be the lipid substrate precursor, and the driving force for the reaction was shown to be a large increase in AS* (7-10). Despite these recent advances in characterizing the reaction and the protein, the relationship of FAEE synthase to any known family of proteins has escaped definition. Our current work not only relates this enzymatic system to detoxification pathways for xenobiotics/carcinogens but also points out its importance to alcohol metabolism in the liver and to cytochrome P450 metabolism of ethanol and carcinogens.

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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Metabolism of ethanol and carcinogens by glutathione transferases.

Bora

Spilburg²,

Lange³

1989

Proc. Natl. Acad. Sci. U.S.A.

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“…Previous studies have described the copurification of FAEE synthase and GST activities from human heart and the sequence similarities between these two enzymes (23,24). GST is known to be abundant in placenta.…”

Section: Mechanism O F Ethanol Teratogenesismentioning

confidence: 99%

Fetal Alcohol Syndrome and Fatty Acid Ethyl Esters

et al. 1992

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ABSTRACT. Fetal alcohol syndrome is the leading known cause of mental retardation. The syndrome, defined as growth retardation, midface hypoplasia, and neurologic dysfunction, represents only part of the spectrum of fetal alcohol effects. The biochemical mechanism of teratogenesis is unknown. In adults, metabolites of ethanol, FAEE, are known to accumulate in major organs. The formation of FAEE is catalyzed by a family of enzymes, FAEE synthases. Our hypothesis is that accumulation of FAEE in the embryo results in fetal alcohol syndrome. We have developed assays for FAEE and FAEE synthase activity using mg of tissue. Using these assays, we have shown the following: Human placenta, mouse placenta, heart, and liver are active in catalyzing the formation of FAEE. One h after maternal ethanol administration on gestational d 14, mouse placenta and fetuses accumulated significant quantities of FAEE. The fatty acid incorporated into FAEE was tissue dependent. Tissues from pregnant animals given ethanol on gestational d 7 showed persistence of FAEE on gestational d 14. We conclude that: 1 ) human and mouse placentas have significant FAEE synthase activity, 2) mouse heart, liver, placenta, and fetal tissues accumulate significant amounts of FAEE after maternal ethanol exposure, 3) there is tissue specificity for the fatty acid incorporated into FAEE, and 4) FAEE may persist for 7 d in placentas. These results provide a basis for further research into the role of FAEE in the development of fetal alcohol syndrome. (Pediatr Res 31: 492-495, 1992)

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“…In addition, another GST3, the major GST isoenzyme expressed in placenta, FAEE synthase isoenzyme purified from human and the GST isoenzyme with the closest N-terminal'semyocardium also displayed GST activity [2]. These quence to that reported for the rnyocardial FAEE synresults suggested that some of the FAEE synthase isoenthase [l].…”

Section: Introductionmentioning

confidence: 83%

Do the major human glutathione S‐transferases have fatty acid ethyl ester synthase activity?

1990

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Two fatty acid ethyl ester (FAEE) synthase isoenzymes purified from human myocardium were reported to be glutathione S-transferases (GST) [(1989) Proc. Natl. Acad. Sci. USA 86, 4470-4473; and (1989) J. Clin. Invest, 84, 1942-19461. In the present study, the FAEE synthase activity of several purified and well characterized human GSTs were examined with ethanol and [14C]oleic acid as substrates. Three isoenzymes, GSTl, GST2 and GST3 which are members of the evolutionary classes /i, CI and II, respectively, were studied and failed to show any significant synthesis of FAEE after 45 min incubation at 37°C. FAEE synthase activity and GS'T3 activity in human placental extracts can be readily separated by ion exchange chromatography on DEAE cellulose, Thus the results show that FAEE synthase activity is not a feature of the major GSTS found in human tissues, The two FAEE synthase isoenzymes isolated by Bora et al. may have been co-purified with GST isoenzymes or these FAEE synthases may be members of the GST super family that have low specific activity in conventional GST assays and have not been previously described.Glutathione S-transferase; Fatty acid ethylester synthase INTRODUCTION In order to determine if FAEE synthase activity is a common feature of all human GSTs or if it is confinedIt has recently been reported [l] that a fatty acid ethyl to particular classes or isoenzymes, we have determined ester (FAEE) synthase isoenzyme purified from human the FAEE synthase activity of several highly purified myocardium had a very similar N-terminal sequence to human GST isoenzymes that represent examples of the that of an acidic human glutathione S-transferase three main evolutionary classes of cytosolic GST. In ad-(GST). The FAEE synthase also had GST activity and dition, we have followed the FAEE synthase activity of VVZ~ able to catalyse the conjugation of GSH and human placental extracts during the purification of 1 -chloro-2,4_dinitrobenzene.In addition, another GST3, the major GST isoenzyme expressed in placenta, FAEE synthase isoenzyme purified from human and the GST isoenzyme with the closest N-terminal'se-

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Identification of a satellite fatty acid ethyl ester synthase from human myocardium as a glutathione S-transferase.

Cited by 36 publications

References 20 publications

Metabolism of ethanol and carcinogens by glutathione transferases.

Metabolism of ethanol and carcinogens by glutathione transferases.

Fetal Alcohol Syndrome and Fatty Acid Ethyl Esters

Do the major human glutathione S‐transferases have fatty acid ethyl ester synthase activity?

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