Dehydratase Inhibition Research Articles

Structural Bases of Dihydroxy Acid Dehydratase Inhibition and Biodesign for Self-Resistance.

Dihydroxy acid dehydratase (DHAD) is the third enzyme in the plant branched-chain amino acid biosynthetic pathway and the target for commercial herbicide development. We have previously reported the discovery of fungal natural product aspterric acid (AA) as a submicromolar inhibitor of DHAD through self-resistance gene directed genome mining. Here, we reveal the mechanism of AA inhibition on DHAD and the self-resistance mechanism of AstD, which is encoded by the self-resistance gene astD. As a competitive inhibitor, the hydroxycarboxylic acid group of AA mimics the binding of the natural substrate of DHAD, while the hydrophobic moiety of AA occupies the substrate entrance cavity. Compared to DHAD, AstD has a relatively narrow substrate channel to prevent AA from binding. Several mutants of DHAD were generated and assayed to validate the self-resistance mechanism and to confer Arabidopsis thaliana DHAD with AA resistance. These results will lead to the engineering of new type of herbicides targeting DHAD and provide direction for the ecological construction of herbicide-resistant crops.

Element Concentrations in Tree Swallows (Tachycineta bicolor) from the U.S. and Binational Great Lakes Areas of Concern.

Selected elements were targeted in state Remedial Action Plans as one group of chemicals affecting the Beneficial Use Impairments of Great Lakes Areas of Concern (AOCs). Livers of nestling tree swallows, Tachycineta bicolor, were harvested from 76 sites in the Great Lakes, which included multiple sites at 27 AOCs and 12 reference sites from 2010 to 2015, and were analyzed for 21 elements. Mercury concentrations were at background levels at all sites. Elevated cadmium (Cd) concentrations were associated with industry. The highest Cd values were from the Black River, OH AOC and were associated with historic coke production but were not at toxic levels. Lead (Pb) concentrations were highest on the Rouge River, MI AOC-the oldest and most heavily populated and industrialized area in southeast Michigan. Individual Pb concentrations were elevated to a level associated with delta-aminolaevulinic acid dehydratase inhibition but not to a level considered toxic. In contrast, livers harvested from sites on the southwest shore of Lake Michigan had selenium (Se) concentrations elevated to levels associated with reduced avian reproduction. One likely source of the high Se concentrations was pollution from a local coal-fired power plant. Concentrations of the remaining elements were at background levels.

Neuroprotective effects of berberine on recognition memory impairment, oxidative stress, and damage to the purinergic system in rats submitted to intracerebroventricular injection of streptozotocin.

Alzheimer's disease (AD) is a progressive and irreversible neurodegenerative disease. The present study investigated the effects of 50 and 100mg/kg berberine (BRB) on recognition memory, oxidative stress, and purinergic neurotransmission, in a model of sporadic dementia of the Alzheimer's type induced by intracerebroventricular (ICV) injection of streptozotocin (STZ) in rats. Rats were submitted to ICV-STZ 3mg/kg or saline, and 3days later, were started on a treatment of BRB or saline for 21days. The results demonstrated that BRB was effective in protecting against memory impairment, increased reactive oxygen species, and the subsequent increase in protein and lipid oxidation in the cerebral cortex and hippocampus, as well as δ-aminolevulinate dehydratase inhibition in the cerebral cortex. Moreover, the decrease in total thiols, and the reduced glutathione and glutathione S-transferase activity in the cerebral cortex and hippocampus of ICV-STZ rats, was prevented by BRB treatment. Besides an antioxidant effect, BRB treatment was capable of preventing decreases in ecto-nucleoside triphosphate diphosphohydrolase (NTPDase), 5'-nucleotidase (EC-5'-Nt), and adenosine deaminase (ADA) activities in synaptosomes of the cerebral cortex and hippocampus. Thus, our data suggest that BRB exerts a neuroprotective effect on recognition memory, as well as on oxidative stress and oxidative stress-related damage, such as dysfunction of the purinergic system. This suggests that BRB may act as a promising multipotent agent for the treatment of AD.

Newborn Screening for Hereditary Tyrosinemia Type I in Québec: Update.

Hereditary tyrosinemia type I (HTI) is a rare autosomal recessive disorder caused by a fumarylacetoacetate hydrolase (FAH) deficiency. If untreated, its acute form is characterized by hepatic failure, renal dysfunction and neurological crisis, and may lead to death. Due to a genetic founder effect in the French-Canadian population, the prevalence of HTI is increased in the province of Quebec (1/19 819), with the IVS12 + 5G>A (1062 + 5G>A) splice site mutation responsible for more than 90% of mutated alleles. Universal newborn screening for (HT1) was thus established in 1970, and close to four million infants have been tested so far, allowing to identify 185 of the 190 affected newborns. During the 1970-1997 period, 2,249,000 newborns were screened at 3-7 days of life on dried filter paper blood spots by tyrosine (Tyr) concentration followed by indirect colorimetric semi-quantitative and quantitative (Q) succinylacetone (SA) testing (red blood cells δ-aminolevulinate dehydratase inhibition), with immunoreactive FAH as the confirmatory test. This approach allowed to identify 118 of 123 affected newborns. In 1998, owing to earlier hospital discharge and increased rate of breastfeeding, four cases were missed within the same year as the discriminating power of blood Tyr became inadequate. Thus, the screening algorithm was modified: indirect semi-quantitative SA measurement became the first-tier test between 1998 and 2014, and direct SA measurement by tandem mass spectrometry (MS/MS) was implemented in 2014, followed by indirect quantitative SA measurement as second tier test. Confirmation is performed by plasmatic amino acid profile and molecular testing. During the 1998-2016 period, more than 1,5 million neonates have been tested (90% sampled between 24 and 48 h of life): 67 of the 67 HTI cases were identified. Both indirect and direct SA measurement as the initial HTI screening test proved to be highly sensitive and specific, with positive and negative predicting value of 79% and 100% respectively.

Nanocurcumin Prevents Oxidative Stress Induced following Arsenic and Fluoride Co-exposure in Rats

<p>The present study is in continuation of our previous efforts to investigate the preventive efficacy of encapsulated curcumin nanoparticles (nanocurcumin) in mitigating effects of arsenic and/or fluoride. Curcumin was encapsulated in chitosan nanoparticles having a size distribution in the range of 50 nm. Sodium arsenite (2 mg/kg, orally) and Fluoride (50 ppm in drinking water) either alone or in combination were administered to male Wistar rats for four weeks to evaluate the efficacy of nanocurcumin (15 mg/kg) in rats. The preventive efficacy of nanocurcumin was evaluated against various altered biochemical variables suggestive of oxidative stress in liver and kidneys, and concentration of As and F in blood. Nanocurcumin co-administration with arsenic and fluoride resulted in lowering of reactive oxygen species and restoration of blood glutathione level which were found to be altered in arsenic and fluoride intoxicated groups. Nanocurcumin were also found to be effective in reversing δ-aminolevulinic acid dehydratase (ALAD) inhibition caused by arsenic exposure. The most promising result from our study shows that nanocurcumin removes not only arsenic but also fluoride from blood which may be due to the enhanced bioavailability and moderate chelating potential of nanocurcumin.</p>

Effects of zinc against mercury toxicity in female rats 12 and 48 hours after HgCl2 exposure.

This work investigated the toxicity of inorganic mercury and zinc preventive effects in female rats sacrificed 12 or 48 h after HgCl2 exposure. Female Wistar rats were subcutaneously injected with ZnCl2 (27 mg/kg) or saline (0.9 %), and 24 h later they were exposed to HgCl2 (5 mg/kg) or saline (0.9 %). Rats sacrificed 12 hours after Hg administration presented an increase in kidney weight and a decrease in renal ascorbic acid levels. Zinc pretreatment prevented the renal weight increase. Rats sacrificed 48 h after Hg exposure presented a decrease in body weight gain, an increase in renal weight, a decrease in renal δ-aminolevulinic acid dehydratase activity, an increase in serum creatinine and urea levels, and a decrease in kidney total thiol levels. Zinc pretreatment partly prevented the decrease in body weight gain and increase in creatinine levels, in addition to totally preventing renal δ-aminolevulinic acid dehydratase inhibition. Mercury accumulation in the kidney and liver in both periods was observed after Hg administration. These results show the different Hg effects along the time of intoxication, and a considerably preventive effect of zinc against Hg toxicity.

Effects of sub-lethal and chronic lead concentrations on blood and liver ALA-D activity and hematological parameters in Nile tilapia

Liver and blood δ-aminolevulinic acid dehydratase (ALA-D) inhibition by exposure to sub-lethal lead concentrations over time in Nile tilapia (Oreochromis niloticus) were investigated. All three lead concentrations (1mgkg−1, 10mgkg−1 and 100mgkg−1) significantly inhibited ALA-D activity in blood (319±29.2; 180±14.6 and 172±19µmols−1h−1L−1 respectively) and liver (302±5.84; 201±41.4 and 93±22.1µmols−1h−1L−1) 24h after injection relative to controls (blood: 597±37.0µmols−1h−1L−1; liver: 376±23.1µmols−1h−1L−1). Blood ALA-D was greatly inhibited in all but the highest lead dose. Fish were then exposed to 1mgkg−1 lead for 9 days, and presented short-term hyperglycemia, decreased hemoglobin and hematocrit values and time-dependent blood ALA-D activity inhibition, corroborating blood ALA-D activity as being more suitable for investigating lead effects, showing dose and time-dependent ALA-D inhibition after lead exposure. The results of the present study also demonstrated that fish size affects blood ALA-D activity, as fish from the 24-h assay, which were slightly smaller (approximately 200g), showed higher ALA-D inhibition in response to lead exposure when compared to the fish from the 9-day assay (approximately 500g). Thus, fish size should always be taken into account both in the field and in laboratory settings, and efforts should be made to obtain uniform fish size samples for biomarker studies.

Are delta-aminolevulinate dehydratase inhibition and metal concentrations additional factors for the age-related cognitive decline?

Aging is often accompanied by cognitive impairments and influenced by oxidative status and chemical imbalances. Thus, this study was conducted to examine whether age-related cognitive deficit is associated with oxidative damage, especially with inhibition of the enzyme delta-aminolevulinate dehydratase (ALA-D), as well as to verify the influence of some metals in the enzyme activity and cognitive performance. Blood ALA-D activity, essential (Fe, Zn, Cu, Se) and non-essential metals (Pb, Cd, Hg, As, Cr, Ni, V) were measured in 50 elderly and 20 healthy young subjects. Cognitive function was assessed by tests from Consortium to Establish a Registry for Alzheimer’s Disease (CERAD) battery and other. The elderly group presented decreased ALA-D activity compared to the young group. The index of ALA-D reactivation was similar to both study groups, but negatively associated with metals. The mean levels of essential metals were within the reference values, while the most toxic metals were above them in both groups. Cognitive function impairments were observed in elderly group and were associated with decreased ALA-D activity, with lower levels of Se and higher levels of toxic metals (Hg and V). Results suggest that the reduced ALA-D activity in elderly can be an additional factor involved in cognitive decline, since its inhibition throughout life could lead to accumulation of the neurotoxic compound ALA. Toxic metals were found to contribute to cognitive decline and also to influence ALA-D reactivation.

Effect of astaxanthin on kidney function impairment and oxidative stress induced by mercuric chloride in rats

Reactive oxygen species are implicated as mediators of tissue damage in the acute renal failure induced by inorganic mercury. Astaxanthin (ASX), a carotenoid with potent antioxidant properties, exists naturally in various plants, algae, and seafoods. This paper evaluated the ability of ASX to prevent HgCl 2 nephrotoxicity. Rats were injected with HgCl 2 (0 or 5 mg/kg b.w., sc) 6 h after ASX had been administered (0, 10, 25, or 50 mg/kg, by gavage) and were killed 12 h after HgCl 2 exposure. Although ASX prevented the increase of lipid and protein oxidation and attenuated histopathological changes caused by HgCl 2 in kidney, it did not prevent creatinine increase in plasma and δ-aminolevulinic acid dehydratase inhibition induced by HgCl 2. Glutathione peroxidase and catalase activities were enhanced, while superoxide dismutase activity was depressed in HgCl 2-treated rats when compared to control and these effects were prevented by ASX. Our results indicate that ASX could have a beneficial role against HgCl 2 toxicity by preventing lipid and protein oxidation, changes in the activity of antioxidant enzymes and histopathological changes.

Delta-aminolevulinic acid dehydratase inhibition and oxidative stress in relation to blood lead among urban adolescents

To explore lead-induced oxidative stress among urban adolescents, the present study, the first from India, was designed to determine the proportion of urban adolescents with blood lead >10 microg/dL and its impact on selected oxidative stress parameters and delta-aminolevulinic acid dehydratase (delta-ALAD) inhibition, which could be used as biomarkers of lead intoxication. A total of 39, urban, male adolescents, drawn from Lucknow and adjoining areas, were recruited to determine lead, delta-ALAD, malondialdehyde (MDA) and glutathione (GSH) in blood and catalase (CAT) in RBCs. Mean level of blood lead was 9.96 +/- 3.63 microg/dL (4.62-18.64); 43% of adolescents crossed the Centre for Disease Control (CDC) intervention level of 10 pg/dL blood lead. On the basis of blood lead levels (BLLs), adolescents were categorized into two groups: Group I and Group II had a blood lead <10 microg/dL (7.40 +/- 1.62) and >10 microg/dL (13.27 +/- 2.67), respectively, with significantly different mean values (P <0.001). Age, sex, body mass index (BMI), Hb level (malnutrition), and area of living as confounders of lead exposure and toxicity were not statistically different between the two groups. However, delta-ALAD activity was significantly lower (P <0.001), while CAT activity was higher in Group II than in Group I (P <0.01). MDA level was also significantly higher in Group II compared to Group I (P <0.001). There were significant negative correlation of BLL with 6-ALAD (r= -0.592, P <0.001), and positive correlations with CAT (r=0.485, P <0.01) and MDA (r=0.717, P <0.001). Interestingly, delta-ALAD, in turn, had significant negative correlations with CAT (r= -0.456, P <0.01) and MDA (r= -0.507, P <0.01). Results of the present pilot study provide clues to the possible low level of lead-induced oxidative stress in urban adolescents, suggesting that lead-induced 6-ALAD inhibition can also be an indicator of oxidative stress. The potential of oxidative stress parameters to be used as biomarkers of lead toxicity warranted further investigation.

Human erythrocyte δ‐aminolevulinate dehydratase inhibition by monosaccharides is not mediated by oxidation of enzyme sulfhydryl groups

The heme pathway enzyme delta-aminolevulinate dehydratase is a good marker for oxidative stress and metal intoxication. This sulfhydryl enzyme is inhibited in such oxidative pathologies as lead, mercury and aluminum intoxication, exposure to selenium organic species and diabetes. Oxidative stress is a complicating factor in diabetes, inducing non-enzymatic glucose-mediated reactions that change protein structures and impair enzyme functions. We have studied the effects of high glucose, fructose and ribose concentrations on delta-ALA-D activity in vitro. These reducing sugars inhibited delta-ALA-D with efficacies in the order fructose=ribose>glucose. The possible mechanism of glucose inhibition was investigated using lysine, DTT, and t-butylamine. Oxidation of the enzyme's critical sulfhydryl groups was not involved because DTT had no effect. We concluded that high concentrations of reducing sugars or their autoxidation products inhibit delta-ALA-D by a mechanism not related to thiol oxidation. Also, we are not able to demonstrate that the formation of a Schiff base with the critical lysine residue of the enzyme is involved in the inhibition of delta-ALA-D by hexoses.

Reversal of aminolevulinic acid dehydratase (ALAD) inhibition and reduction of erythrocyte protoporphyrin levels by Vitamin C in occupational lead exposure in Abeokuta, Nigeria

In order to investigate the toxic effects of lead during occupational exposure to this metal, the activity of aminolevulinic acid dehydratase (ALAD) and the concentrations of erythrocyte protoporphyrin (EPP) were determined in blood of various artisans in Abeokuta, Nigeria and these were related to blood lead levels. ALAD activity in the artisans was inhibited to varying extents. ALAD activity was inhibited to the tune of 77% in petrol station attendants while the lowest inhibition of 36% was obtained in the welders. EPP was also significantly increased in the artisans ( p < 0.001). The highest EPP level of 241.57 ± 89.27 μg/100 ml of red blood cell was observed in upholsterers. A significant ( p < 0.001) negative correlation was observed between ALAD activity and blood lead levels on one hand ( r = −0.631) and between ALAD activity and EPP on the other hand ( r = −0.461). Administration of a daily dose of 500 mg ascorbic acid for 2 weeks reversed the lead-induced inhibition of ALAD. Increased EPP levels observed in the artisans also responded positively to the ascorbic acid supplementation. A significant reduction ( p < 0.001) in blood lead was also observed in the artisans at the end of the 2-week ascorbic acid therapy. Our findings indicate that ascorbic acid may offer more advantages over the conventional agents for the treatment of lead poisoning, especially in cases where the subjects cannot be removed from the source of lead exposure.

Ceruloplasmin Oxidase Activity as a Biomarker of Lead Exposure

The most sensitive method used in monitoring the blood lead-level is atomic absorption spectrophotometry. Its limitation due to its high cost has made delta-aminolaevulinic acid dehydratase (delta-ALAD) inhibition more popular. This work demonstrates that ceruloplasmin (Cp), the blue-colored copper containing alpha-2 globulin (alpha(2) globulin) glycoprotein in serum, can serve as a high-potential biomarker for lead exposure. Sephadex G-200 column chromatography demonstrated that lead-intoxicated patient serum transferrin and ceruloplasmin are bound to lead. The ceruloplasmin's oxidase and delta-ALAD activities are reduced when the blood-lead level increases (thresholds of 10.0 and 12.5 mug/100 mL respectively), with a correlation coefficient of -0.8643 and -0.6785 which accurately estimate a blood lead level of 74.7% and 46.0%, respectively. When the blood-lead level was less than 20 mug/100 mL, correlation coefficients of -0.5785 and -0.6963 suggested that Cp may serve as a good biomarker. Other advantages in using Cp in monitoring the blood-lead level were its stability: the enzyme was more stable at -20 degrees C, for more than 2 wk, and its practical and rapid determination procedure.

Mechanism of delta-aminolevulinate dehydratase inhibition by phenyl selenoacetylene involves its conversion to diphenyl diselenide

The mechanism of delta-aminolevulinate dehydratase (delta-ALA-D) inhibition by phenyl selenoacetylene in vitro was investigated in this study. Phenyl selenoacetylene (40-400 microM) inhibition of delta-aminolevulinate dehydratase from rat liver (low speed supernatant fraction, S1 fraction) was partially prevented by incubation under argon atmosphere and completely prevented by dithiothreitol. After incubation with S1 fraction from rat liver or cysteine (40 mM), phenyl selenoacetylene was partially converted into diphenyl diselenide, which is a stronger inhibitor of delta-aminolevulinate dehydratase than phenyl selenoacetylene. Diphenyl diselenide increased the rate of oxidation of -SH groups, while phenyl selenoacetylene did not affect such oxidation. delta-Aminolevulinate dehydratase purified from bovine liver (Sigma) was less sensitive to phenyl selenoacetylene and diphenyl diselenide than the enzyme from S1 fraction. We propose that the lower sensitivity of purified enzyme to selenides could be related to the formation of selenols due to the presence of dithiothreitol (a reducing agent) in the incubation medium. In agreement, incubation of purified enzyme (Sigma) with diphenyl diselenide (2 microM) and sodium borohydride (a reducing agent) under argon atmosphere significantly increased enzyme activity. Results obtained suggest that delta-aminolevulinate dehydratase inhibition by phenyl selenoacetylene is dependent on its conversion into diphenyl diselenide that induces oxidation of essential -SH groups of delta-aminolevulinate dehydratase. We propose that oxygen could be important in the regeneration of diphenyl diselenide leading to a catalytic oxidation of the enzyme.

Erythrocyte aminolevulinic acid dehydratase inhibition by cis-platin

The effect of cis-platin on erythrocyte aminolevulinic acid dehydratase (ALAD) activity was studied in vivo and in vitro. Young male Wistar rats were treated with a single i.p. injection of cis-platin at 2.5, 5.0, and 10.0 mg/kg dose. In addition, a single i.p. injection of lead nitrate (1.0 mg/kg dose) was administered as positive control. Experiments in vitro were also performed to elucidate the possible mechanism of action. The aminolevulinic acid dehydratase was almost completely inhibited in vitro from 0.5 mM concentration, and the IC 50 was stated at 0.265 mM, 20 times higher than lead (IC 50 stated at 0.013 mM). Reduced glutathione, partially but significantly, reactivated in vitro the enzyme treated with cis-platin (0.5 and 5.0 mM), whereas zinc showed a positive, significant effect with the higher dose (5.0 mM) only. On the contrary, inhibition caused by lead (0.005 mM) was partially, but significantly restored by reduced glutathione, and, almost completely, by zinc. The experiments in vivo show that cis-platin causes a dose- and time-dependent inhibition of ALAD activity with 5.0 and 10.0 mg/kg dose, until 66 and 33% of the control activity 96 h after treatment, respectively. The results show that erythrocyte ALAD is sensitive to cis-platin and suggest that the mechanism of enzyme inhibition is a direct interaction with sulfhydryl groups, whereas zinc site appears involved with the higher doses only. This mechanism appears different from lead that prevalently inhibits ALAD removing zinc from the enzyme, other than interacting with sulfhydryl groups.

Experimental lead poisoning in turkey vultures (Cathartes aura).

Lead-induced mortality appears to have been a major factor in the decline of the California condor (Gymnogyps californianus). We orally dosed turkey vultures (Cathartes aura) with BB-sized lead shot from January 1988 through July 1988 to determine physiologic response (delta-aminolevulinic acid dehydratase inhibition, erythrocyte protoporphyrin levels, anemia), diagnostic tissue lead concentrations (blood, liver, and kidney), and comparative sensitivity of this species. Two turkey vultures died and two became so intoxicated they were euthanized. Overall, responses of measured parameters were comparable to other species exposed to lead although there was considerable individual variation. Survival time (143-211 days), even with the large numbers of shot and constant redosing, was much longer than reported for other species of birds, suggesting considerable tolerance by turkey vultures to the deleterious effects of lead ingestion. Based on these observations, turkey vultures appear to be poor models for assessing the risk of lead poisoning to California condors or predicting their physiologic response.

Delta-aminolevulinate dehydratase inhibition by phenyl selenoacetylene: effect of reaction with hydrogen peroxide.

The effect of phenyl selenoacetylene and its selenoxide on delta-aminolevulinate dehydratase from liver of adult rats (mammalian source) and from cucumber leaves (plant source) was investigated. In vivo, selenides can be oxidized to selenoxides by flavin-containing monooxygenases and selenoxides can regenerate selenides by thiol oxidation. The compound phenyl selenoacetylene was converted to selenoxides by reaction with hydrogen peroxide. Phenyl selenoacetylene inhibited mammalian and plant delta-aminolevulinate dehydratase with an IC50 about 250 microM and >400 microM, respectively. Its selenoxide inhibited the enzyme more strongly, with IC50 values of 45 microM and 100 microM for the mammalian and plant source, respectively. The selenoxide inhibitory action was antagonized by dithiothreitol suggesting the involvement of -SH groups. Moreover, delta-aminolevulinate dehydratase from a plant source was inhibited by the selenoxide, suggesting a possible involvement of -SH groups located at a site distinct from the region implicated in Zn2+ binding in mammalian delta-aminolevulinate dehydratase. The results of the present study suggest that (i) delta-aminolevulinate dehydratase is a potential molecular target for phenyl selenoacetylene, due to the oxidation of enzyme sulfhydryl groups, and that (ii) the monooxygenation of this selenocompound, which in vivo could be possibly mediated by flavin-containing monooxigenases, increases its inhibitory effect.

Delta-Aminolevulinate dehydratase inhibition by 2,3-dimercaptopropanol is mediated by chelation of zinc from a site involved in maintaining cysteinyl residues in a reduced state.

The mechanisms underlying mouse delta-aminolevulinate dehydratase (ALA-D) inhibition by a chelating agent used in the treatment of heavy metal poisoning, 2,3-dimercaptopropanol (British Anti-Lewisite), were investigated. ALA-D inhibition by 2,3-dimercaptopropanol was totally reversed by 25-100 microM Zn2+, indicating that inhibition was due to chelation of zinc by 2,3-dimercaptopropanol. Our data suggested that zinc bound to a labile site (displaced by 25-40 microM EDTA or 500 microM 2,3-dimercaptopropanol) is involved in maintaining the sulfhydryl groups of ALA-D in a reduced state (essential for enzyme activity), since inhibition by these compounds was reversed by 10 mM dithiotreitol (a reducing agent). On the other hand, 10 mM dithiotreitol did not reverse ALA-D inhibition by a higher concentration of EDTA (100 microM). Accordingly, 2,3-dimercaptopropanol appears to inhibit ALA-D through a mechanism similar to that of low EDTA concentrations. Neither oxidized 2,3-dimercaptopropanol nor reactive oxygen species appeared to contribute for ALA-D inhibition by reduced 2,3-dimercaptopropanol. Taken together, these results suggest that 2,3-dimercaptopropanol inhibits ALA-D by chelating Zn2+ from a labile site that is involved in maintaining enzyme sulfhydryl groups in a reduced state. This site is compatible with the ZnB or Zn beta previously described in mammalian and bacterial ALA-D.

A CASE OF TYROSINAEMIA TYPE I WITH NORMAL LEVEL OF SUCCINYLACETONE IN THE AMNIOTIC FLUID

Prenatal diagnosis of tyrosinaemia type I can be achieved in cultured amniotic cells and in chorionic villus material by testing the activity of fumarylacetoacetate hydrolase and by DNA analysis, and in amniotic fluid by succinylacetone measurement. This specific metabolite can be measured either by gas chromatography-mass spectrometry or by delta-aminolevulinate dehydratase inhibition assay. In a series of 65 at-risk cases tested with the enzyme inhibition assay, one case out of the 18 with the disease had a normal level of succinylacetone. This case is presented.

An acquired acute hepatic porphyria: A novel type of δ-aminolevulinate dehydratase inhibition

An acquired acute hepatic porphyria: A novel type of δ-aminolevulinate dehydratase inhibition