Red Cell Enzyme Activities Research Articles

“Lysosomal” enzyme activities in red blood cells of normal individuals and patients with homozygous β-thalassaemia

Four hydrolases, β-galactosidase, β-glucuronidase, β- N-acetylglucosaminidase and acid phosphatase were examined in red blood cells (RBC) of normal donors and patients with homozygous ß-thalassaemia. Highly sensitive fluorimetric substrates were used to determine the specific activities of these enzymes. In order to avoid contamination by lysosomal activities derived from white blood cells (WBC), the mature RBC were separated from other blood elements by cellulose chromatography. The hydrolase activities in normal RBC were detected only in their plasma membranes and were found to be considerably lower than in WBC or platelets. In thalassaemic RBC, hydrolase activities were present in both plasma membranes and in the soluble fraction. The normoblast fraction contributed most of the hydrolase activity found in these preparations, suggesting the presence of lysosomal particles in thalassaemic RBC. No differences in the enzymatic activities were found when purified membranes of mature RBC from thalassemic and normal preparations were compared. The origin and roles of these hydrolytic enzymes in normal and thalassaemic RBC membranes are not known.

Red blood cell enzyme abnormalities in patients treated with chemotherapy.

Sxiteen red blood cell enzyme activities and fetal haemoglobin level have been assayed in 60 patients treated for haematologic or nonhaematologic malignant diseases with various combinations of cytostatic drugs. Acquired enzyme deficiency was found in 20 patients. The most frequently decreased activities were those of G6PD (12 cases), PK (seven cases), PFK (six cases) and AK (three cases). In many patients decreased activity of some enzymes contrasted with increased activity of others such as ALD, TPI, G3PD, PGK, ENOL and 6PGD. The number of abnormalities seems to be related to the duration of the treatment. Incidence of enzyme deficiencies was similar in patients previously treated or not with radiotherapy. Enzyme abnormalities were correlated neither with haemoglobin level nor with reticulocyte count. They were very similar to those observed in primary acquired dyserythropoietic and preleukaemic states. Their prognostic value and mechanism are discussed.

Red cell enzyme activities and properties of mutant pyruvate kinase after long-term storage of red cells in liquid nitrogen.

After 8 months of storage of normal red cells in liquid nitrogen, red cell enzyme activities were essentially unchanged, except for triose phosphate isomerase which was reduced by about half. Mutant pyruvate kinase (PK), PK Tokyo I, showed no changes in kinetic or electrophoretic properties after 8 months of storage. These findings indicate that red cells stored for a long term in liquid nitrogen can still be used for the study of mutant PK.

Enzymes of galactose metabolism in erythrocytes and liver of inbred strains of mice

This study compares the activity of galactokinase, galactose-1-phosphate uridyltransferase, and UDPgalactose-4-epimerase, the important enzymes in the pathway of conversion of galactose to glucose in red cells and liver of five inbred strains of mice. In the red cells, galactokinase varied over a four-fold range of activity while the other enzymes varied about two-fold. The activity of each of the enzymes varied independently of the other so that red cells of each strain had a unique pattern for the three enzymes. The red cell activity pattern was not reflected in liver tissue which showed little interstrain variation for each of the three enzymes. The ratio of liver galactokinase to uridyltransferase and epimerase was very similar in all five strains. Oxidation in vivo of 14C galactose to 14CO 2 was examined in the two strains of mice with the widest divergence of red cell galactokinase activity and no difference was found in this parameter measuring the physiological disposition of the sugar. The wide variation of the red cell enzyme activity appears to have little metabolic consequence for the animal, the oxidation of the sugar reflecting the relative constancy of the liver enzyme activity.

Significance of the determination of red cell enzyme activities.

American Journal of HematologyVolume 6, Issue 2 p. 163-172 Concise Reviews Significance of the determination of red cell enzyme activities Professor Shiro Miwa MD, Corresponding Author Professor Shiro Miwa MD Third Department of Internal Medicine, Yamaguchi University School of Medicine, Ube, JapanThird Department of Internal Medicine, Yamaguchi University School of Medicine, 1144 Kogushi, Ube-shi, Yamaguchi-ken 755, JapanSearch for more papers by this author Professor Shiro Miwa MD, Corresponding Author Professor Shiro Miwa MD Third Department of Internal Medicine, Yamaguchi University School of Medicine, Ube, JapanThird Department of Internal Medicine, Yamaguchi University School of Medicine, 1144 Kogushi, Ube-shi, Yamaguchi-ken 755, JapanSearch for more papers by this author First published: April 1979 https://doi.org/10.1002/ajh.2830060209Citations: 5AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume6, Issue2April 1979Pages 163-172 RelatedInformation

Hexokinase Deficiency in Erythrocytes: A New Variant in 5 Members of a Finnish Family

15 cases of congenital haemolytic anaemia have thus far been attributed to hexokinase (HK) deficiency in erythrocytes. We report some clinical, biochemical and genetic findings from 5 members of a Finnish family with this deficiency. The proband, a 1-year-old girl, was the only patient with anaemia. All subjects had either mild or marked reticulocytosis. Red cell ATP levels were at the lower range of normal in all subjects and 2,3-DPG was abnormally low in one. The activities of red cell enzymes, other than HK, were within or above the normal range, respectively. The Km-values for glucose and fructose were elevated (ATP normal) in the subjects with HK deficiency. We speculate that the family represents heterozygosity of a mutant allele and that there is phenotypic variation associated with the HK mutant. The locus might be subject to mutations which lead to a variety of HK variants and to a spectrum of diseases. This point of view is in accordance with the overwhelming variation of reaction kinetics and metabolic effects of this and other reported cases.

Red Cell Enzyme Activities and Properties of Mutant Pyruvate Kinase after Long-Term Storage of Red Cells in Liquid Nitrogen

Red Cell Enzyme Activities and Properties of Mutant Pyruvate Kinase after Long-Term Storage of Red Cells in Liquid Nitrogen

Erythrocyte enzymes in groups of Rattus norvegicus with genetic differences in 2,3-diphosphoglycerate levels

1. 1. A major locus with two alleles is responsible for large differences in erythrocyte 2,3-diphosphoglycerate (DPG) levels in Rattus norvegicus. Blood from homozygous High-DPG, homozygous Low-DPG and heterozygous animals was used to measure blood indices and red cell enzyme activities. 2. 2. Significant differences between groups were found in DPG levels, white blood cell counts and hemoglobin levels. 3. 3. The results suggest that none of the red cell enzymes assayed is structurally or quantitatively different in the three groups.

The effects of splenectomy on the hematology and on the activity of delta-aminolevulinic acid dehydratase (ALA-D) in hematopoietic tissues of the european eel, Anguilla anguilla L.

1. 1. The effects of splenectomy on hematological parameters and on the activity of ALA-D in some tissues of the European eel, Anguilla anguilla L., were studied after 1, 2, 4, 6 and 13 weeks. 2. 2. No marked differences in red blood cell variables (hematocrit, hemoglobin and RBC count) or enzyme activity were observed between sham-operated and splenectomized eels. 3. 3. An initial lymphopenia followed later by neutrophilia were observed in both operated groups. 4. 4. Abnormal erythrocytes were present in the blood from splenectomized eels. 5. 5. The spleen seems to function primarily as an erythrocyte reservoir and destruction organ for erythrocytes and has only a subsidiary role as an erythrocyte-producing organ. Lymphocytopoiesis seems to occur partly in the spleen, while granulocytopoiesis probably occurs elsewhere.

Heterogeneity of erythrocyte pyruvate kinase deficiency and related metabolic disorders in patients with hematological diseases

In several patients suffering from congenital non-spherocytic hemolytic anemia or from malignant hematological disorder associated with erythrocyte pyruvate kinase (PK) deficiency, a metabolic study has been carried out involving the following biochemical determinations: assay of red cell enzyme activities; estimation of glucose consumption; measurement of the rate of glycolytic intermediates; and, in some cases, enzyme purification and characterization of the PK variant. Metabolic equilibrium most probably does not depend on kinetic characteristics of PK molecules. Furthermore, the data obtained allow separation of cases with congenital non-spherocytic hemolytic anemia (hereditary defect) and acquired PK deficiencies.

Deficiency of adenosine deaminase not associated with severe combined immunodeficiency

The 12-year-old Kung (""Bushman'') boy from South West Africa who has marked deficiency of red cell adenosine deaminase has been found to have 2 to 3% of enzyme activity in red blood cells, 10 to 12% in leukocytes, and 10 to 30% in cultured fibroblasts. The enzyme has ADA 1 electrophoretic mobility: SV40 transformation of cultured fibroblasts caused a decrease of ""tissue ADA'' and an increase in ""red cell ADA'' isozymes. A battery of investigations revealed that the child has normal humoral and cellular immunity. A family study showed that a sibling had the same level of red cell ADA and the parents had intermediate levels. Studies of the Kung population from which the child comes have shown that the allele responsible for the condition, and which we designate ADA8, is polymorphic.

Action of Growth Hormone on Erythropoiesis: Changes in Red Blood Cell Enzyme Activities in Growth-retarded Patients with and without Growth Hormone Deficiency

Fifteen red cell enzyme activities of growth-retarded patients with and without growth hormone (GH) deficiency were investigated before and after GH administration. The 15 enzymes were Hexokinase, phosphoglucomutase, glucose phosphate, isomerase, phosphofructokinase, fructose diphosphate aldolase, glyceraldehyde-3-phosphae dehydrogenase, triosephosphate isomerase, 2,3-diphosphoglycerate mutase, 3-phosphoglycerate kinase, 3-phosphoglycerate mutase, enolase, pyruvate kinase, glycose-6-phosphate dehydrogenase, 6-phosphogluconic dehydrogenase, glutathione reducase. Sixty-six subjects were studied: 30 normal control subjects (group N) and 36 patients (aged 5-23 years) with short stature. Complete endocrine evaluation showed 21 (group I) to have GH deficiency (10 patients with isolated GH deficiency) and 15 (group II) to have normal hypothalamic and pituitary function except for two patients with a moderate hypothyroidism. Both had been receiving thyroid hormone treatment for a long time before our studies. All 36 patients were treated with 2 mg human growth hormone intramuscularly for 7 days. Before GH treatment no significant difference was observed between hematologic data in group I (GH deficiency) and group II (no GH deficiency). After GH therapy there was a significant increase in reticulocyte count in both groups of patients with short stature. The mean pretreatment value in group I was 1.294% +/- 0.084 (SEM); the mean post-treatment value was 2.081% +/- 0.287 (SEM)< P less than 0.005. The mean pretreatment value in group II was 1.0% 0.184 (SEM); the mean post-treatment value was 1.407% +/- 0.193 (SEM), P less than 0.01. In group II (no GH deficiency) mean pretreatment erythrocyte enzyme activities were not significantly different from those activities observed in normal control subjects (group N). However, in patients who lacked GH, the pretreatment activities of five red cell enzymes (glucose phosphate isomerase, triosephosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, 2,3-diphosphoglycerate mutase, 3-phosphoglycerate kinase) were significantly decreased before GH administration compared with the values in normal control subjects...

Glucose-6-phosphate dehydrogenase Ferrara. A new variant of g-6-PD identified in Northern Italy.

A new variant of glucose-6-phosphate dehydrogenase (G-6-PD) has been discovered in Northern Italy, in the district of Ferrara. This variant is characterized by high decrease of red blood cell enzyme activity (less than 5% of normal), high affinity for G-6-P and NADP, increased utilization of deamino-NADP and 2-deoxy-G-6-P, and faster electrophoretic mobility in the buffer systems commonly used for the classification of the G-6-PD variants. The new G-6-PD type was never associated with clinical manifestations in any cases except neonatal jaundice in some of the newborns with this enzyme deficiency. The frequency of the new variant in the Ferrara district indicates that it has probably appeared in this area by mutation some centuries ago. It is suggested that this variant should be named G-6-PD Ferrara.

Ethnic Variation in Red Cell Glutathione Peroxidase Activity

Glutathione peroxidase activity was measured in blood and cultured fibroblasts from healthy persons of several different population groups. Individuals of Jewish ancestry and others of Mediterranean origin were found to manifest a decrease of red cell but not of leukocyte or fibroblast enzyme activity. Oriental populations differed in that the scatter in red cell enzyme activity was significantly lower than in Occidental populations. The erythrocyte enzyme of individuals with low activity was found to be less stable to heating than was the enzyme from persons with high activity. As a possible explanation for these data, a provisional genetic model is presented: a low GSH Px allele with a frequency of 0.556 in the Jewish population and of only 0.181 in the United States-Northern European population. Our results suggest that an association between GSH Px deficiency and hemolytic anemia need not represent a cause-and-effect relationship.

Ethnic variation in red cell glutathione peroxidase activity

Glutathione peroxidase activity was measured in blood and cultured fibroblasts from healthy persons of several different population groups. Individuals of Jewish ancestry and others of Mediterranean origin were found to manifest a decrease of red cell but not of leukocyte or fibroblast enzyme activity. Oriental populations differed in that the scatter in red cell enzyme activity was significantly lower than in Occidental populations. The erythrocyte enzyme of individuals with low activity was found to be less stable to heating than was the enzyme from persons with high activity. As a possible explanation for these data, a provisional genetic model is presented: a low GSH Px allele with a frequency of 0.556 in the Jewish population and of only 0.181 in the United States-Northern European population. Our results suggest that an association between GSH Px deficiency and hemolytic anemia need not represent a cause-and-effect relationship.

Acute infantile hemiplegia with cerebrospinal fluid eosinophilic pleocytosis: An unusual case of visceral larva migrans

1. Deiss A, Lee GR, and Cartwright GE: Hemolytic anemia in Wilson's disease, Ann Intern Med 73:413, 1970. 2. Melntyre N, Clink HM, Levi AJ, Cumings JN, and Sherlock S: Hemolytic anemia in Wilson's disease, N Engl J Med 276:439, 1967. 3. Slovis TL, Dubois RS, Rodgerson DO, and Silverman A: The varied manifestations of Wilson's disease, J PEDIATR 78:578, 1971. 4. Walshe JM: Wilson's disease. The presenting sYmptoms, Arch Dis Child 37:253, 1962. 5. Deiss A, Lynch RE, Lee GR, and Cartwright GE: Longterm therapy of Wilson's disease, Ann Intern Med 75:57, 1971. 6. Oski F: Chickee, the copper, Ann Intern Med 73:485, 1970. 7, Cartwright GE, Hodges RE, Gubler CJ, Mahoney JP, Daum K, Wintrobe MM, and Bean WB: Studies on copper metabolism. XIII. Hepatolenticular degeneration, J Clin Invest 33:1487, 1954. 8. Boulard M, Blume L-G, and Beutler E: The effect of copper on red cell enzyme activities, J Clin Invest 51:459, 1972. 9. Feig SA, Segal GB, Shohet SB, and Nathan DG: Energy metabolism in human erythrocytes. II. Effects of glucose depletion, J Clin Invest 51:1547, 1972.

A longitudinal study of red cell enzymes in infants of low birth weight.

A longitudinal study of red cell enzyme activity in newborn infants of low birth weight has been conducted over the first 2 months of life. The enzymes investigated are acetylcholinesterase (EC 3.1.3.7), an integral part of the red cell membrane and subnormal in ABO hemolytic disease of the newborn; and glucose-6-phosphate dehydrogenase (EC 1.1.1.49), an intracellularly-located, sex-linked enzyme, implicated in neonatal jaundice and of significance in drug-induced hemolytic anemias. Acetylcholinesterase activity, which is lower in normal full-term infants and in low birth weight infants than in adults, was further diminished during the initial weeks of life of the infants of low birth weight and the higher levels of activity, characteristic of adult red cells, had not appeared by 2 months of age. By contrast, red cell glucose-6-phosphate dehydrogenase activity, which is higher in full-term newborn infants and in infants of low birth weight than in adults, did not diminish as a function of age and the lower adult levels were not discernible by 2 months of life.

A prolidase deficiency in man with iminopeptiduria

A deficiency in the enzyme, prolidase (EC 3.4.3.7), is described in a patient with iminopeptiduria and clinical findings that suggest a defect in collagen metabolism. The massive iminopeptiduria is unique to this disorder, with daily proline excretion rates of 1.76 g. Fifteen peptides were characterized and quantitated from a deproteinized, amino-acid-free, 24-hr urine collection using column chromatographic procedures. Amino acid analysis revealed the peptides to be di- and tri-peptides containing proline plus another amino acid, “X.” Dansylation revealed “X” to be amino-terminal in each case. Prolidase, an enzyme known to cleave X-proline bonds, was measured by two separate methods. Enzyme activity in the patient's red and white cells was absent to markedly decreased as compared to age-matched controls. Furthermore, the activity of the enzyme in the mother's and maternal grandfather's white cells was less than that found in adult controls, suggesting a heterozygous condition. The maternal grandmother's enzyme activity was normal. The enzyme data is suggestive of an autosomal recessive disorder. This patient represents the first direct demonstration of a prolidase deficiency in man.

A new method for separation of human blood components.

MOST of the enzyme activities in red cells are lower than those in white cells. To examine the enzyme activities and minor components in red cells, the red cells must first be separated from the white cells. The repeated centrifugations in various media usually used for this purpose are time-consuming and there is also the disadvantage of an accompanying loss of the youngest population of red cells, which frequently show stronger enzyme activities than older red cells1. When such preparations are used, the enzyme activities may thus be underestimated. This causes difficulty in interpreting data, especially when specimens from different individuals are compared. Filtrations, for example, through cotton and glass wool are incomplete in separation2.

A dimethyltryptamine-forming enzyme in human blood.

An enzyme capable of forming the hallucinogen dimethyltryptamine was found in human red blood cells, plasma, and platelets. The enzyme activity in red blood cells and plasma was not significantly different in psychiatric patients from that in normal subjects. The enzyme activity in platelets was higher in psychotic subjects than in nonpsychotics and was apparently related to the presence of a dialyzable inhibitor in the normal subjects.