ALA-dehydratase activity inZea mays L.

Peroxidase (EC 1.11.1.7) enzyme units that were recovered by sequential extraction at low and high ionic strength buffer pH 7 from a peanut suspension culture at various intervals of cell growth appeared to increase with culture time. In particular, at the end of a 14-day growth cycle, peroxidase specific activity rose while protein content and the specific activity of porphobilinogen synthase (aminolevulinic acid, dehydratase EC 4.2.1.24) declined. The decrease of the protein content could probably be accounted for by the 95% decrease of protein synthesis at the end of the growth phase. The highest specific activity of aminolevulinic acid dehydratase (ALA dehydratase) appeared to be associated with organelles which could be precipitated at 500 × g and this specific activity appeared to be unaffected by the stage of growth of these cells. It is suggested that the ALA dehydratase activity in nonchlorophyllous peanut cells, which is as high as that found in green tissue, may play a function in the biosynthesis of the hemoprotein peroxidase.

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delta-Aminolevulinate dehydratase in human erythroleukemia cells: an immunologically distinct enzyme

Blood ◽

10.1182/blood.v65.4.939.939 ◽

1985 ◽

Vol 65 (4) ◽

pp. 939-944 ◽

Cited By ~ 13

Author(s):

CS Chang ◽

S Sassa

Keyword(s):

Butyric Acid ◽

Deae Cellulose ◽

K562 Cells ◽

Normal Adult ◽

Erythroleukemia Cells ◽

Normal Enzyme ◽

Normal Human ◽

Dehydratase Activity ◽

Ala Dehydratase ◽

Selenium Oxide

Abstract Physicochemical and immunologic properties of delta-aminolevulinate (ALA) dehydratase in human K562 erythroleukemia cells were examined. ALA dehydratase activity was found to increase in K562 cells after treatment with butyric acid or selenium oxide. Enzyme activity in untreated K562 cells was comparable to that in normal adult erythrocytes but was increased three- to six-fold in K562 cells treated with 1.2 mmol/L butyric acid or 0.03 mmol/L selenium oxide. The Michaelis-Menten constant (Km), the inhibitor constant (Ki), and elution profile by diethylaminoethyl (DEAE) cellulose chromatography were similar for ALA dehydratase from K562 cells and normal human adult and human fetal erythrocytes. However, ALA dehydratase from K562 cells did not react with a monospecific rabbit antibody against ALA dehydratase purified from normal adult erythrocytes, although the antibody reacted with the enzyme from normal adult and fetal red cells. These findings indicate that ALA dehydratase in K562 cells is immunologically distinct from the normal enzyme.

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Erythrocyte δ-aminolaevulinic Acid Dehydratase Activity and Blood Protoporphyrin Concentrations as Indices of Lead Exposure and Altered Haem Biosynthesis

Clinical Science ◽

10.1042/cs0560061 ◽

1979 ◽

Vol 56 (1) ◽

pp. 61-69 ◽

Cited By ~ 14

Author(s):

P. A. Meredith ◽

M. R. Moore ◽

A. Goldberg

Keyword(s):

Lead Exposure ◽

Lead Concentration ◽

False Negative ◽

Blood Lead ◽

Blood Lead Concentration ◽

Least Squares Regression ◽

Exponential Relationship ◽

Aminolaevulinic Acid ◽

Dehydratase Activity ◽

Ala Dehydratase

1. The activity of erythrocyte δ-aminolaevulinic acid (ALA) dehydratase and blood protoporphyrin concentrations have been measured in patients with various anaemias, a group of subjects with known lead exposure and a group of control subjects. Leucocyte ALA synthase was measured in subjects from the last two groups. 2. Erythrocyte ALA dehydratase activity was significantly depressed in the group of lead-exposed subjects and showed a highly significant negative exponential relationship with blood lead concentration. 3. Blood protoporphyrin concentrations were significantly elevated in the group of lead-exposed subjects and patients with iron-deficiency anaemia and showed a significant positive exponential relationship with blood lead concentration. 4. Comparison of the least-squares regression analysis of these relationships and incidence of false positive and false negative results indicates that erythrocyte ALA dehydratase activity is a more accurate measure of environmental and moderate industrial lead exposure than blood protoporphyrin concentrations. 5. The correlations of erythrocyte ALA dehydratase and leucocyte ALA synthase activity, and of blood protoporphyrin concentrations and leucocyte ALA synthase activity, suggest that blood protoporphyrin more accurately reflects haem synthesis than does erythrocyte ALA dehydratase activity.

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Induction of delta-aminolevulinic acid dehydratase in mouse Friend virus-transformed erythroleukemia cells during erythroid differentiation

Blood ◽

10.1182/blood.v64.1.64.64 ◽

1984 ◽

Vol 64 (1) ◽

pp. 64-70 ◽

Cited By ~ 15

Author(s):

CS Chang ◽

S Sassa

Keyword(s):

Normal Mouse ◽

De Novo ◽

Aminolevulinic Acid ◽

Erythroid Differentiation ◽

Friend Virus ◽

Enzyme Protein ◽

Direct Demonstration ◽

Murine Erythroleukemia ◽

Dehydratase Activity ◽

Ala Dehydratase

Abstract The activity of delta-aminolevulinic acid (ALA) dehydratase, an enzyme involved in heme biosynthesis, has been shown to increase in Friend virus-transformed murine erythroleukemia (MEL) cells during erythroid differentiation. In this study, the nature of the increase in ALA dehydratase activity in MEL cells was examined using a monospecific antibody directed to the enzyme. A sevenfold increase in ALA dehydratase activity was observed after cells had been treated with 1.5% Me2SO for 5 days. Ouchterlony double immunodiffusion analysis showed that lysates from untreated and Me2SO-treated MEL cells formed a single precipitin line with rabbit IgG directed to the normal mouse liver ALA dehydratase. A single arc of identity was also observed with the lysates from normal mouse erythrocytes, spleen, liver, and lysates from both uninduced and induced MEL cells. Rocket immunoelectrophoresis demonstrated that lysates from both uninduced and induced cells formed rockets with the IgG and that the peak height of the rocket was proportional to the ALA dehydratase activity applied. The slope of linear plots of rocket peak heights v ALA dehydratase activity was identical for lysates from uninduced and Me2SO-induced cells. Succinylacetone, a potent inhibitor of ALA dehydratase, was shown to markedly inhibit the activity of the enzyme, but did not interfere with the synthesis of ALA dehydratase induced by Me2SO treatment. Me2SO- induced increases in ALA dehydratase activity and the enzyme protein were both blocked by the simultaneous treatment of cells with 5-bromo- 2′-deoxyuridine (BrdU). BrdU-mediated repression of ALA dehydratase was partially overcome by treating the cells with thymidine. These data demonstrate that increased ALA dehydratase activity in MEL cells undergoing erythroid differentiation after Me2SO treatment is due to de novo synthesis of the same enzyme protein present in uninduced MEL cells as well as in normal erythrocytes. This represents the first direct demonstration of an increase in a heme biosynthetic pathway enzyme protein in erythroid cells undergoing differentiation.

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delta-Aminolevulinate dehydratase in human erythroleukemia cells: an immunologically distinct enzyme

Blood ◽

10.1182/blood.v65.4.939.bloodjournal654939 ◽

1985 ◽

Vol 65 (4) ◽

pp. 939-944

Author(s):

CS Chang ◽

S Sassa

Keyword(s):

Butyric Acid ◽

Deae Cellulose ◽

K562 Cells ◽

Normal Adult ◽

Erythroleukemia Cells ◽

Normal Enzyme ◽

Normal Human ◽

Dehydratase Activity ◽

Ala Dehydratase ◽

Selenium Oxide

Physicochemical and immunologic properties of delta-aminolevulinate (ALA) dehydratase in human K562 erythroleukemia cells were examined. ALA dehydratase activity was found to increase in K562 cells after treatment with butyric acid or selenium oxide. Enzyme activity in untreated K562 cells was comparable to that in normal adult erythrocytes but was increased three- to six-fold in K562 cells treated with 1.2 mmol/L butyric acid or 0.03 mmol/L selenium oxide. The Michaelis-Menten constant (Km), the inhibitor constant (Ki), and elution profile by diethylaminoethyl (DEAE) cellulose chromatography were similar for ALA dehydratase from K562 cells and normal human adult and human fetal erythrocytes. However, ALA dehydratase from K562 cells did not react with a monospecific rabbit antibody against ALA dehydratase purified from normal adult erythrocytes, although the antibody reacted with the enzyme from normal adult and fetal red cells. These findings indicate that ALA dehydratase in K562 cells is immunologically distinct from the normal enzyme.

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Depression of Erythrocyte δ-Aminolaevulinic Acid Dehydratase Activity in Alcoholics

Clinical Science ◽

10.1042/cs0460415 ◽

1974 ◽

Vol 46 (3) ◽

pp. 415-418 ◽

Cited By ~ 3

Author(s):

N. Krasner ◽

M. R. Moore ◽

G. G. Thompson ◽

W. McIntosh ◽

A. Goldberg

Keyword(s):

Alcohol Consumption ◽

Alcohol Withdrawal ◽

Leucine Aminopeptidase ◽

Blood Alcohol ◽

Alcohol Content ◽

Blood Alcohol Content ◽

Aminolaevulinic Acid ◽

Glutamyl Transpeptidase ◽

Dehydratase Activity ◽

Ala Dehydratase

1. δ-Aminolaevulinic acid (ALA) dehydratase, γ-glutamyl transpeptidase (γGT) and leucine aminopeptidase (LAP) have been measured in chronic alcoholics. 2. Within 48 h of alcohol withdrawal ALA dehydratase activity was depressed in chronic alcoholics even though the blood alcohol content was zero. A second group of chronic alcoholics studied 7 days after the cessation of alcohol consumption had normal ALA dehydratase activities. 3. ALA dehydratase activity is more reliable than measurements of γGT and LAP in monitoring alcohol habits of chronic alcoholics.

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Effect of some metals and related metal-orgaic compounds on ALA-dehydratase activity of corn

Plant and Soil ◽

10.1007/bf02182145 ◽

1984 ◽

Vol 79 (1) ◽

pp. 69-75 ◽

Cited By ~ 7

Author(s):

L. Scarponi ◽

P. Perucci

Keyword(s):

Dehydratase Activity ◽

Ala Dehydratase

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Induction of delta-aminolevulinic acid dehydratase in mouse Friend virus-transformed erythroleukemia cells during erythroid differentiation

Blood ◽

10.1182/blood.v64.1.64.bloodjournal64164 ◽

1984 ◽

Vol 64 (1) ◽

pp. 64-70

Author(s):

CS Chang ◽

S Sassa

Keyword(s):

Normal Mouse ◽

De Novo ◽

Aminolevulinic Acid ◽

Erythroid Differentiation ◽

Friend Virus ◽

Enzyme Protein ◽

Direct Demonstration ◽

Murine Erythroleukemia ◽

Dehydratase Activity ◽

Ala Dehydratase

The activity of delta-aminolevulinic acid (ALA) dehydratase, an enzyme involved in heme biosynthesis, has been shown to increase in Friend virus-transformed murine erythroleukemia (MEL) cells during erythroid differentiation. In this study, the nature of the increase in ALA dehydratase activity in MEL cells was examined using a monospecific antibody directed to the enzyme. A sevenfold increase in ALA dehydratase activity was observed after cells had been treated with 1.5% Me2SO for 5 days. Ouchterlony double immunodiffusion analysis showed that lysates from untreated and Me2SO-treated MEL cells formed a single precipitin line with rabbit IgG directed to the normal mouse liver ALA dehydratase. A single arc of identity was also observed with the lysates from normal mouse erythrocytes, spleen, liver, and lysates from both uninduced and induced MEL cells. Rocket immunoelectrophoresis demonstrated that lysates from both uninduced and induced cells formed rockets with the IgG and that the peak height of the rocket was proportional to the ALA dehydratase activity applied. The slope of linear plots of rocket peak heights v ALA dehydratase activity was identical for lysates from uninduced and Me2SO-induced cells. Succinylacetone, a potent inhibitor of ALA dehydratase, was shown to markedly inhibit the activity of the enzyme, but did not interfere with the synthesis of ALA dehydratase induced by Me2SO treatment. Me2SO- induced increases in ALA dehydratase activity and the enzyme protein were both blocked by the simultaneous treatment of cells with 5-bromo- 2′-deoxyuridine (BrdU). BrdU-mediated repression of ALA dehydratase was partially overcome by treating the cells with thymidine. These data demonstrate that increased ALA dehydratase activity in MEL cells undergoing erythroid differentiation after Me2SO treatment is due to de novo synthesis of the same enzyme protein present in uninduced MEL cells as well as in normal erythrocytes. This represents the first direct demonstration of an increase in a heme biosynthetic pathway enzyme protein in erythroid cells undergoing differentiation.

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Fetal Effects of Lead Exposure

PEDIATRICS ◽

10.1542/peds.49.1.145a ◽

1972 ◽

Vol 49 (1) ◽

pp. 145-146

Author(s):

John Scanlon

Keyword(s):

Lead Exposure ◽

Whole Blood ◽

Aminolevulinic Acid ◽

Suckling Rats ◽

Average Child ◽

Lead Absorption ◽

Lead Levels ◽

Dehydratase Activity ◽

Ala Dehydratase

Dr. Chisolm's commentary on increased lead absorption in September's Pediatrics was thoughtful and timely. One part of the lead picture which needs a great deal of further study is the fetal effects of exposure to increasing amounts of lead from the environment. It has been demonstrated recently that concentrations of lead found in the "average" child (20 to 40 µg/100 gm whole blood) can inhibit Δ-aminolevulinic acid (ALA) dehydratase activity. This same article points out that in suckling rats these "usual" lead levels inhibit ALA dehydratase activity in both blood and brain.

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