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Showing papers on "Aldehyde dehydrogenase published in 1985"


Journal ArticleDOI
TL;DR: The 500-residue amino acid sequence of the subunit of mitochondrial human liver aldehyde dehydrogenase is reported, the first structure determined for this enzyme type from any species, and is based on peptides from treatments with trypsin, CNBr, staphylococcal Glu-specific protease, and hydroxylamine.
Abstract: The primary structure of the mitochondrial form of horse liver aldehyde dehydrogenase has been determined, utilizing peptide analyses and homology with other enzyme forms. The subunit exhibits N-terminal heterogeneity in size similar to that for the corresponding human mitochondrial protein, the longest form having 500 residues. Catalase was identified as a contaminant of the preparations. All four pairs within a set of aldehyde dehydrogenases can now be compared, including the same two species variants (horse and human) for both the cytosolic and mitochondrial enzyme, revealing characteristic differences although Cys-302 and other segments of presumed functional importance are unchanged. The cytosolic and mitochondrial enzymes are clearly different (172 exchanges in the horse pair; 160 exchanges in the human pair) and the mitochondrial forms are more conserved (28 exchanges of 500 residues) than the cytosolic ones (43 exchanges). Distributions of the residue substitutions also differ between the two enzyme types. These results suggest a comparatively distant separation of the cytosolic and mitochondrial enzymes into forms with separate functional constraints that are more strict on the mitochondrial than the cytosolic enzyme. Unexpectedly, positions with residues unique to one of the four enzymes are about twice as common in both of the horse proteins than in either of the human proteins. This difference may reflect a general pattern for human/non-human proteins, showing that not only functional properties of the protein, but also other factors, such as generation time (longer in man than in horse), are important for enzyme divergence.

147 citations


Journal ArticleDOI
TL;DR: Dietary ethanol inhibited FAS and repressed the accumulation of TG in ADH-deficient larvae, suggesting that the levels of these factors may be subject to a complex feedback control.
Abstract: When cultured on a defined diet, ethanol was an efficient substrate for lipid synthesis in wild-type Drosophila melanogaster larvae. At certain dietary levels both ethanol and sucrose could displace the other as a lipid substrate. In wild-type larvae more than 90% of the flux from ethanol to lipid was metabolized via the alcohol dehydrogenase (ADH) system. The ADH and aldehyde dehydrogenase activities of ADH were modulated in tandem by dietary ethanol, suggesting that ADH provided substrate for lipogenesis by degrading ethanol to acetaldehyde and then to acetic acid. The tissue activity of catalase was suppressed by dietary ethanol, implying that catalase was not a major factor in ethanol metabolism in larvae. The activities of lipogenic enzymes, sn-glycerol-3-phosphate dehydrogenase, fatty acid synthetase (FAS), and ADH, together with the triacylglycerol (TG) content of wild-type larvae increased in proportion to the dietary ethanol concentration to 4.5% (v/v). Dietary ethanol inhibited FAS and repressed the accumulation of TG in ADH-deficient larvae, suggesting that the levels of these factors may be subject to a complex feedback control.

120 citations


Journal ArticleDOI
TL;DR: Isoelectric focusing and electrophoresis were used to identify the various isozymes of alcohol dehydrogenase (ADH), aldehyde dehydrogenases (ALDH), aldhyde oxidase (AOX), and xanthine oxidases (XOX) and the physiological role of each enzyme is discussed.
Abstract: Isoelectric focusing and electrophoresis were used to identify the various isozymes of alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH), aldehyde oxidase (AOX), and xanthine oxidase (XOX). ADH types I, II, and III were located primarily in the cytosol fraction of liver, but some activity was found also in the small granule fraction. The ALDH-I and -IV isozymes were found in the large granule fraction, while ALDH-II and -III were present in the cytosol and ALDH-V in the small granule fraction. AOX and XOX each appeared as a single cytosolic form with some small granule activity. The tissue distribution of these isozymes is presented and the physiological role of each enzyme is discussed.

100 citations


Journal Article
TL;DR: Observations strongly suggest that aldehyde dehydrogenase activity is an important determinant with regard to the sensitivity of a cell population to the oxazaphosphorines.
Abstract: The sensitivity of cultured L1210 and P388 cells sensitive (L1210/0, P388/0) and resistant (L1210/OAP, P388/CLA) to oxazaphosphorines, to 4-hydroperoxycyclophosphamide, ASTA Z-7557, phosphoramide mustard, and acrolein was determined in the absence and presence of known (disulfiram, diethyldithiocarbamate, cyanamide) or suspected [ethylphenyl(2-formylethyl)phosphinate] inhibitors of aldehyde dehydrogenase activity. The L1210/OAP cell line is resistant specifically to the oxazaphosphorines; P388/CLA cells are partially cross-resistant to other cross-linking agents. All four inhibitors of aldehyde dehydrogenase activity potentiated the cytotoxic action of the oxazaphosphorines, 4-hydroperoxycyclophosphamide and ASTA Z-7557, against L1210/OAP and P388/CLA cells; in the presence of a sufficient amount of inhibitor, sensitivity was essentially fully restored in both cases. The inhibitors did not potentiate the cytotoxic action of the nonoxazaphosphorines, phosphoramide mustard and acrolein, against these cell lines. The cytotoxic action of the oxazaphosphorines and nonoxazaphosphorines against L1210/0 and P388/0 cells was not potentiated by any of the aldehyde dehydrogenase inhibitors. Inhibitors of xanthine oxidase or aldehyde oxidase activities did not potentiate the cytotoxic action of the oxazaphosphorines against L1210/OAP cells. These observations strongly suggest that (a) aldehyde dehydrogenase activity is an important determinant with regard to the sensitivity of a cell population to the oxazaphosphorines; (b) L1210/0 and P388/0 cells lack the relevant aldehyde dehydrogenase activity; (c) the phenotypic basis for the resistance to oxazaphosphorines by L1210/OAP cells is aldehyde dehydrogenase activity; and (d) the major reason that P388/CLA cells are resistant to oxazaphosphorines is aldehyde dehydrogenase activity.

96 citations


Journal ArticleDOI
TL;DR: Ex vivo sensitivity of murine pluripotent hematopoietic stem cells and myeloid progenitor cells to 4-hydroperoxycyclophosphamide, ASTA Z 7557, phosphoramide mustard, acrolein, melphalan, and cis-platinum was determined and the phenotypic basis for the relative insensitivity of CFU-S to oxazaphosphorines is the aldehyde dehydrogenase activity contained by these cells

73 citations


Journal ArticleDOI
TL;DR: Measurements of oxygen consumption revealed that the electron transport system was capable of reoxidizing ALDH-generated NADH much faster than it was produced and hence was not rate-limiting for aldehyde metabolism.

70 citations


Journal ArticleDOI
01 May 1985-Alcohol
TL;DR: Individuals deficient in aldehyde dehydrogenase isozyme may consume less alcohol, as per capita alcohol consumption correlated with the frequency of isozyme deficiency and blood acetaldehyde level after alcohol drinking was found significantly higher in deficient subjects than in individuals without deficiency.

67 citations


Journal ArticleDOI
TL;DR: Comparison of kinetic constants for aldehydes among the enzymes indicated that alcohol dehydrogenase is the best reductase with the highest affinity and Kcat values.

63 citations


Journal ArticleDOI
TL;DR: The biochemical properties of ALDH isozymes have been examined in human tissues and one set, designated ALDH3, has been studied in detail, and high levels of expression were found in human‐rodent hybrids, constructed using rat hepatoma cells, and these hybrids were used to assign the human AL DH3 gene to chromosome 17.
Abstract: Summary The biochemical properties of ALDH isozymes have been examined in human tissues and one set, designated ALDH3, has been studied in detail. These components occur at highest levels in lung and stomach, but were not expressed in fetal tissues, or in blood, hair roots and fibroblasts. The ALDH3 isozymes show optimal activity with benzaldehyde and can use either NAD or NADP as cofactor. Antiserum against a partially purified ALDH3, from stomach, selectively precipitates this isozyme from human tissues and selectively recognizes an homologous component in the rat. Human and rodent ALDH3 were not immunoprecipitated by anti-ALDH1 or anti-ALDH2 antisera. High levels of expression were found in human-rodent hybrids, constructed using rat hepatoma cells, and these hybrids were used to assign the human ALDH3 gene to chromosome 17.

55 citations



Journal ArticleDOI
TL;DR: The results show that ADH and ALDH phenotypes among American Indians living in New Mexico are very similar to those of Caucasian populations and quite different from those of Orientals.
Abstract: Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) isoenzyme phenotypes were determined in autopsy liver samples from 50 North American Indians from New Mexico. Forty-six of the 50 livers had sufficient ADH activity to allow phenotyping at the ADH2 and ADH3 loci. All 46 livers possessed the "typical" ADH2 1-1 phenotype. The frequency of the ADH3(2) allele was 0.59 and is the highest thus far reported in any racial population. All 50 livers possessed the ALDH I isoenzyme which exhibits the greatest anodic mobility on starch gel electrophoresis at pH 7.6. The results show that ADH and ALDH phenotypes among American Indians living in New Mexico are very similar to those of Caucasian populations and quite different from those of Orientals.

Journal ArticleDOI
01 Jan 1985-Alcohol
TL;DR: Although H2O2 inhibited yeast aldehyde dehydrogenase in vitro and cyanamide inhibited hepatic catalase in vivo, the possible in hepatic H 2O2 concentration following cyanamide administration does not account for the effects of cyanamide on ethanol metabolism.

Journal ArticleDOI
TL;DR: It is found that the nitrate ester antianginal drugs, isosorbide dinitrate and nitroglycerin, are potent inhibitors of human erythrocyte aldehyde dehydrogenase, and these therapies have well-documented disulfiram-like side effects.
Abstract: Disulfiram-like responses to various drug therapies are caused by elevated ethnol-derived blood acetaldehyde concentrations resulting from drug-induced inhibition of aldehyde dehydrogenase enzymes. We have found that the nitrate ester antianginal drugs, isosorbide dinitrate and nitroglycerin, are potent inhibitors of human erythrocyte aldehyde dehydrogenase. To further characterize this drug-induced enzyme inhibition, erythrocyte aldehyde dehydrogenase activities were measured in patients undergoing therapy with nitrate ester antianginals (isosorbide dinitrate and nitroglycerin) and sulfonylurea hypoglycemics (chlorpropamide and tolazamide). The erythrocyte enzyme was decreased by approximately 25% in sulfonylurea-treated patients, whereas in the nitrate ester-treated patients, an 88% inhibition was observed. The minimal enzyme inhibition in the sulfonylurea-treated group was unexpected because these therapies have well-documented disulfiram-like side effects. This weak inhibition contrasted with the severe inhibition observed in the nitrate ester-treated group where the disulfiram-like side effects are not considered a serious clinical problem. This apparent anomaly was attributed to differences in inhibition of the erythrocyte and liver aldehyde dehydrogenase by the parent drugs and their hepatic metabolites.

Journal ArticleDOI
01 May 1985-Alcohol
TL;DR: Population genetic studies on the prevalence of aldehyde dehydrogenase isozyme I deficiency in various Caucasian, Oriental, African, and American Indian subjects were carried out using hair roots as peripheral source of the enzyme activity, suggesting an autosomal codominant mode of inheritance.

Journal ArticleDOI
TL;DR: The role of alcohol-sensitizing drugs as pharmacological and psychological deterrents, in the treatment of alcoholism, and the action of three new hepatic aldehyde dehydrogenase inhibitors, including methyltetrazolethiol, which maintain that this type of inhibitor could represent a significant advance in the adjunctive pharmacotherapy of alcoholism.

Book ChapterDOI
01 Jan 1985
TL;DR: Most of the acetaldehyde formed from ethanol is subsequently oxidized to acetate in liver, and under normal conditions it is oxidized further so rapidly that significant acetaldehyde concentrations can only be found in the liver.
Abstract: Acetaldehyde is the first oxidation product of ethanol, and under normal conditions it is oxidized further so rapidly that significant acetaldehyde concentrations can only be found in the liver. Aldehyde oxidase, xanthine oxidases, and aldehyde dehydrogenases are all capable of catalyzing aldehyde oxidation. The first two enzymes, however, have a broad substrate specificity and a low affinity for acetaldehyde (K m > 1 mM), and consequently their involvement in the metabolism of acetaldehyde is insignificant (Lundquist 1970; Lindros 1978). The main enzyme oxidizing acetaldehyde is aldehyde dehydrogenase (ALDH), which catalyzes the oxidation of acetaldehyde in the presence of nicotinamide-adenine dinucleotide (NAD) as follows: $$ C{H_3}CHO + NA{D^ + }\xrightarrow[{{H_2}O}]{{ALDH}}C{H_3}CO{O^ - } + NADH + {H^ + }. $$ Most of the acetaldehyde formed from ethanol is subsequently oxidized to acetate in liver.

Journal ArticleDOI
01 May 1985-Alcohol
TL;DR: In the experiment of the repeated ingestion of ethanol in the deficient group, the second peak of blood acetaldehyde level was lower than that of the first one, and beta 60 and ER were not clearly elevated with the increase of ethanol dose, while those in the normal ALDH group increased depending on the blood ethanol level.

Journal ArticleDOI
TL;DR: The specific activity of ADH-FF was 2–3 times higher thanADH-SS with both ethanol and acetaldehyde as dehydrogenase substrates, and ADH and ALDH activities were inhibited by pyrazole, disulphiram and p -hydroxymercuribenzoate.
Abstract: 1. 1. Alcohol dehydrogenase (ADH) allozymes (FF, SF, SS) were purified to homogeneity from strains of Drosophila melanogaster using a new procedure. 2. 2. All allozymes displayed aldehyde dehydrogenase activity on cellulose (ALDH) acetate zymograms, corresponding to the ADH activity zones. 3. 3. Kinetic analyses with acetaldehyde as substrate revealed non-linear, biphasic Lineweaver-Burk plots giving two apparent Michaelis constants ( K m ) ranges for the allozymes of 1–10 μM and 0.1–1 mM. 4. 4. In contrast, kinetic analyses with ethanol as substrate gave results consistent with a single K m value of approximately 2 mM. 5. 5. At approx. 3 μM substrate concentration, the enzymes exhibited equivalent rates of dehydrogenase activity with either ethanol or acetaldehyde; whereas at a concentration of 10 mM, the ADHs exhibited approx. a 10-fold higher activity with ethanol as substrate. 6. 6. The specific activity of ADH-FF was 2–3 times higher than ADH-SS with both ethanol and acetaldehyde as dehydrogenase substrates. ADH and ALDH activities were inhibited by pyrazole, disulphiram and p -hydroxymercuribenzoate. 7. 7. Atomic absorption spectrometry confirmed the absence of zinc. 8. 8. The oxidation of 2-[ 13 C]-labelled ethanol by ADH allozymes in vitro was studied using nuclear magnetic resonance spectrometry. 9. 9. Acetaldehyde, its diol and acetate were detected within 20 min and monitored for 10 hr. 10. 10. The significance of these results for studies on ethanol metabolism in D. melanogaster is discussed.

Journal ArticleDOI
TL;DR: A derivative and possible physiological metabolite of disulfiram, diethyldithiocarbamic acid methanethiol mixed disulfide, is shown here for the first time to inactivate the mitochondrial low‐K m, isozyme of human aldehyde dehydrogenase.

Journal ArticleDOI
TL;DR: Results indicate that aldehyde dehydrogenase is located in the peroxisomal membrane, similar to a clofibrate-induced microsomal enzyme.
Abstract: A study was made of the effect of chronic administration of the hypolipidemic drug clofibrate on the activity and intracellular localization of rat liver aldehyde dehydrogenase. The enzyme was assayed using several aliphatic and aromatic aldehydes. Clofibrate treatment caused a 1.5 to 2.3-fold increase in the liver specific aldehyde dehydrogenase activity. The induced enzyme has a high Km for acetaldehyde and was found to be located in peroxisomes and microsomes. Clofibrate did not alter the enzyme activity in the cytoplasmic fraction. The total peroxisomal aldehyde dehydrogenase activity increased 3 to 4-fold under the action of clofibrate. Disruption of the purified peroxisomes by the hypotonic treatment or in the alkaline conditions resulted in the release of catalase from the broken organelles, while aldehyde dehydrogenase as well as nucleoid-bound urate oxidase and the peroxisomal membrane marker NADH:cytochrome c reductase remained in the peroxisomal ‘ghosts’. At the same time, treatment by Triton X-100 led to solubilization of the membrane-bound NADH:cytochrome c reductase and aldehyde dehydrogenase from intact peroxisomes and their ‘ghosts’. These results indicate that aldehyde dehydrogenase is located in the peroxisomal membrane. The peroxisomal aldehyde dehydrogenase is active with different aliphatic and aromatic aldehydes, except for formaldehyde and glyceraldehyde. The enzyme Km values lie in the millimolar range for acetaldehyde, propionaldehyde, benzaldehyde and phenylacetaldehyde and in the micromolar range for nonanal. Both NAD and NADP serve as coenzymes for the enzyme. Aldehyde dehydrogenase was inhibited by disulfiram, N-ethylmaleimide and 5,5′-dithiobis(2-nitrobenzoic)acid. According to its basic kinetic properties peroxisomal aldehyde dehydrogenase seems to be similar to a clofibrate-induced microsomal enzyme. The functional role of both enzymes in the liver cells is discussed.

Journal ArticleDOI
TL;DR: Hepatic aldehyde dehydrogenase activity is depressed in alcoholic liver disease and may account for the observation that alcoholics develop high blood acetaldehyde concentrations following ethanol, and activity in Group I was significantly lower than in Groups II or III.

Journal ArticleDOI
01 Jan 1985-Alcohol
TL;DR: The subcellular distributions of aldehyde dehydrogenase activities towards acetaldehyde have been determined in wedge-biopsy samples of human liver and the activity in the cytosol was more sensitive to inhibition by disulfiram and had alkaline pH optima.

Journal ArticleDOI
TL;DR: This study provides genetic and physiological evidence for the role of fatty aldehyde as an essential metabolic intermediate and NADP-dependent FALDH as a key enzyme in the dissimilation of hexadecane, hexadecanol, and dodecyl alde Hyde in Acinetobactor sp.
Abstract: The role of fatty aldehyde dehydrogenases (FALDHs) in hexadecane and hexadecanol metabolism was studied in Acinetobacter sp. strain HO1-N. Two distinct FALDHs were demonstrated in Acinetobacter sp. strain HO1-N: a membrane-bound, NADP-dependent FALDH activity induced 5-, 15-, and 9-fold by growth on hexadecanol, dodecyl aldehyde, and hexadecane, respectively, and a constitutive, NAD-dependent, membrane-localized FALDH. The NADP-dependent FALDH exhibited apparent Km and Vmax values for decyl aldehyde of 5.0, 13.0, 18.0, and 18.3 microM and 537.0, 500.0, 25.0, and 38.0 nmol/min in hexadecane-, hexadecanol-, ethanol-, palmitate-grown cells, respectively. FALDH isozymes ald-a, ald-b, and ald-c were demonstrated by gel electrophoresis in extracts of hexadecane- and hexadecanol-grown cells. ald-a, ald-b, and ald-d were present in dodecyl aldehyde-grown cells, while palmitate-grown control cells contained ald-b and ald-d. Dodecyl aldehyde-negative mutants were isolated and grouped into two phenotypic classes based on growth: class 1 mutants were hexadecane and hexadecanol negative and class 2 mutants were hexadecane and hexadecanol positive. Specific activity of NADP-dependent FALDH in Ald21 (class 1 mutant) was 85% lower than that of wild-type FALDH, while the specific activity of Ald24 (class 2 mutant) was 55% greater than that of wild-type FALDH. Ald21R, a dodecyl aldehyde-positive revertant able to grow on hexadecane, hexadecanol, and dodecyl aldehyde, exhibited a 100% increase in the specific activity of the NADP-dependent FALDH. The oxidation of [3H]hexadecane byAld21 yielded the accumulation of 61% more fatty aldehyde than the wild type, while Ald24 accumulated 27% more fatty aldehyde, 95% more fatty alcohol, and 65% more wax ester than the wild type. This study provides genetic and physiological evidence for the role of fatty aldehyde as an essential metabolic intermediate and NADP-dependent FALDH as a key enzyme in the dissimilation of hexadecane, hexadecanol, and dodecyl aldehyde in Acinetobactor sp. strain HO1-N. Images

Journal ArticleDOI
TL;DR: It is suggested that AHD-5 may be the primary enzyme for oxidizing mitochondrial acetaldehyde during ethanol oxidation in vivo, and may also exist as a monomeric form as well.

Journal ArticleDOI
TL;DR: It is suggested that nonenzymatic reactions between indole‐3‐acetaldehyde (or other biogenic aldehydes and membrane components might occur in vivo, and could be involved in the effects of drugs such as ethanol and barbiturates.
Abstract: When indole-3-acetaldehyde was incubated with rat brain tissue, an aldehyde dehydrogenase-independent irreversible disappearance of the aldehyde was found. This was accompanied by an increase in absorbance at 240–400 nm, with a peak at 310 nm. The results suggested that this change in absorbance was caused by a membrane-bound nonenzymatic reaction between indole-3-acetaldehyde and phospholipids. A similar reaction occurred between indole-3-acetaldehyde and pure preparations of phosphatidylethanolamine and phosphatidylserine, but not phosphatidylcholine. Indole-3-acetaldehyde levels also decreased slightly when incubated with albumin but absorbance at 310 nm was unaltered. It is suggested that nonenzymatic reactions between indole-3-acetaldehyde (or other biogenic aldehydes) and membrane components might occur in vivo, and could be involved in the effects of drugs such as ethanol and barbiturates.

Journal ArticleDOI
TL;DR: The coupled alcohol dehydrogen enzyme/aldehyde dehydrogenase has been extended to preparatory scale synthesis of optically pure l -α-hydroxyacids in the presence of a cofactor regeneration system and the active-site cubic-space section model has been shown not to be applicable to all substrates.

Journal ArticleDOI
TL;DR: The above data argue against a significant role of acetaldehyde in the in vivo response of testosterone to ethanol, and appear to impair gonadotropin-testicular receptor interaction in vivo.

Journal ArticleDOI
TL;DR: According to substrate specificity, kinetic and molecular properties clofibrate-induced aldehyde dehydrogenase appears to be identical to normal liver microsomal enzyme.

Journal ArticleDOI
TL;DR: The in vitro inactivation of aldehyde dehydrogenase by cyanamide in rat liver slices, in intact mitochondria, and at various stages of purity was characterized, suggesting an alternative pathway for ALDH inactivation may exist in which enzymatic modification of cyanamide is not necessary.

Journal ArticleDOI
TL;DR: Aldehyde reductases (alcohol: NADP+-oxidoreductase, EC 1.1.2) I and II from human placenta have been purified to homogeneity and exhibit strong affinity towards various other aldehydes such as glyceraldehyde, propionaldehyd, and pyridine-3-aldehyde.