Molecular basis of glutathione reductase deficiency in human blood cells

Blood ◽  
2006 ◽  
Vol 109 (8) ◽  
pp. 3560-3566 ◽  
Author(s):  
Nanne M. Kamerbeek ◽  
Rob van Zwieten ◽  
Martin de Boer ◽  
Gert Morren ◽  
Herma Vuil ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Glutathione Reductase ◽  
Blood Cells ◽  
Stop Codon ◽  
Residual Activity ◽  
Premature Stop Codon ◽  
Dimerization Domain ◽  
Human Blood Cells ◽  
Glutathione Reductase Deficiency ◽  
Conserved Residue

Abstract Hereditary glutathione reductase (GR) deficiency was found in only 2 cases when testing more than 15 000 blood samples. We have investigated the blood cells of 2 patients (1a and 1b) in a previously described family suffering from favism and cataract and of a novel patient (2) presenting with severe neonatal jaundice. Red blood cells and leukocytes of the patients in family 1 did not contain any GR activity, and the GR protein was undetectable by Western blotting. Owing to a 2246-bp deletion in the patients' DNA, translated GR is expected to lack almost the complete dimerization domain, which results in unstable and inactive enzyme. The red blood cells from patient 2 did not exhibit GR activity either, but the patient's leukocytes contained some residual activity that correlated with a weak protein expression. Patient 2 was found to be a compound heterozygote, with a premature stop codon on one allele and a substitution of glycine 330, a highly conserved residue in the superfamily of NAD(P)H-dependent disulfide reductases, into alanine on the other allele. Studies on recombinant GR G330A revealed a drastically impaired thermostability of the protein. This is the first identification of mutations in the GR gene causing clinical GR deficiency.

THE EFFECT OF NECROGENIC DIETS ON THE ACTIVITIES OF CERTAIN ENZYME SYSTEMS IN THE RED BLOOD CELLS OF RATS

1964 ◽  
Vol 42 (12) ◽  
pp. 1809-1814 ◽  
Author(s):  
Beverley B. Lazier ◽  
J. M. R. Beveridge
Keyword(s):  
Red Blood Cells ◽  
Glutathione Reductase ◽  
Dehydrogenase Activity ◽  
Blood Cells ◽  
Sodium Selenite ◽  
Basal Diet ◽  
Hepatic Necrosis ◽  
Male Rats ◽  
Enzyme Systems ◽  
Glucose 6 Phosphate Dehydrogenase

The enzymic activities of glucose-6-phosphate dehydrogenase, glutathione reductase, and catalase were studied in the red blood cells of male rats which were fed a basal diet designed to induce acute hepatic necrosis. Significant decreases in the activity were found for all three systems. These changes were prevented by supplementing the basal diet with one of the following: methionine, sodium selenite, DL-α-tocopherol acetate, or all three factors plus cystine.There was no significant change in 6-phosphogluconic dehydrogenase activity when it was investigated under similar circumstances.

Antioxidant Status of Red Blood Cells and Liver in Hypercholesterolemic Rats Fed Hypolipidemic Spices

2004 ◽  
Vol 74 (3) ◽  
pp. 199-208 ◽  
Author(s):  
Kempaiah ◽  
Srinivasan
Keyword(s):  
Antioxidant Enzymes ◽  
Red Blood Cells ◽  
Glutathione Reductase ◽  
Blood Cells ◽  
Antioxidant Status ◽  
Animal Studies ◽  
Glutathione Transferase ◽  
Reductase Activity ◽  
Glutathione S Transferase ◽  
Dietary Spices

Animal studies were carried out to examine the beneficial influence of known hypolipidemic spice principles – curcumin, capsaicin, and garlic – on the antioxidant status of red blood cells and liver under induced hypercholesterolemic conditions. Groups of experimental rats rendered hypercholesterolemic were maintained on curcumin (0.2%)/capsaicin (0.015%)/garlic (2.0% dry powder)-containing diets for eight weeks. Erythrocytes isolated at the end of the study were analyzed for intracellular antioxidant molecules and antioxidant enzymes. Intracellular thiols and glutathione content in red blood cells were significantly depleted (by about 35%) in hypercholesterolemic rats. This depletion in intracellular thiols and glutathione was effectively countered by dietary spice principles – curcumin, capsaicin, and garlic. Glutathione reductase activity that was lowered in hypercholesterolemic conditions (by 25%) was completely countered by dietary spice principles and garlic. Activities of glutathione transferase, glutathione peroxidase, and catalase in erythrocytes remained unchanged under hypercholesterolemic conditions. Although hemoglobin levels of erythrocytes were not affected, methemoglobin concentration was significantly increased in hypercholesterolemic rats. This alteration was partially countered by dietary spice principles. Significant fall in hepatic total thiols in the hypercholesterolemic situation was partially corrected by dietary spice treatment. Similarly, the lowered activities of hepatic antioxidant enzymes – glutathione reductase, glutathione-S-transferase, catalase, and superoxide dismutase – in hypercholesterolemic rats were effectively countered by the dietary spices treatment.

scholarly journals Familial deficiency of glutathione reductase in human blood cells

Blood ◽  
1976 ◽  
Vol 48 (1) ◽  
pp. 53-62 ◽  
Author(s):  
H Loos ◽  
D Roos ◽  
R Weening ◽  
J Houwerzijl
Keyword(s):  
Glutathione Reductase ◽  
Blood Cells ◽  
Normal Values ◽  
Reductase Activity ◽  
White Blood Cells ◽  
Human Blood Cells ◽  
Glutathione Reductase Activity ◽  
Fava Beans

A virtually complete absence of glutathione reductase activity was found in the erythrocytes of all three children (one male, two females) from a consanguineous marriage. Intermediate values were found in the erythrocytes of both parents. The enzyme activity could not be restored either by addition of FAD in vitro or by administration of riboflavin in vivo. The amount of reduced glutathione in the erythrocytes was normal in each case. Severely diminished glutathione stability during incubation with acetylphenylhydrazine was observed in the erythrocytes of the siblings, as well as intermediate stability in the parents' red cells. Clinically, this deficiency was manifested by hemolytic crises after eating fava beans in the eldest daughter (patient), and possibly by cataracts in her own and in her brother's eyes. Very low activities of glutathione reductase were also found in the leukocytes of this family: 13%-15% of normal values for the children and 64%-66% for the parents. Moreover, the same deficiency was found in the purified white blood cells of the propositus: 8% of normal values in the polymorphonuclear (PMN) cells, 4% in the lymphocytes, and 15% in the monocytes, together with 11% in the platelets. Finally, we found an abnormal oxygen consumption of the propositus' PMNs after phagocytosis of zymosan particles, suggesting that the glutathione reductase reaction was involved in the bactericidal capacity of these cells.

scholarly journals Molecular Cloning and Characterization of Decay-Accelerating Factor Deficiency in Cromer Blood Group Inab Phenotype

Blood ◽  
1998 ◽  
Vol 91 (2) ◽  
pp. 680-684
Author(s):  
Li Wang ◽  
Makoto Uchikawa ◽  
Hatsue Tsuneyama ◽  
Katsushi Tokunaga ◽  
Kenji Tadokoro ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Group ◽  
Blood Cells ◽  
Stop Codon ◽  
Japanese Woman ◽  
Blood Group System ◽  
Decay Accelerating Factor ◽  
Exon 2 ◽  
Reading Frame ◽  
Old Japanese

An additional decay-accelerating factor (DAF) mutation, designated as Inab phenotype in the Cromer blood group system, was recently identified in a 28-year-old Japanese woman (H.A.). The red blood cells of H.A., like those of other Inab phenotype individuals, were negative for Cromer system antigens, Cra, Tca, Dra, UMC, and IFC. The deficiency of DAF on the red blood cells of H.A. has been shown by immunoblotting with a murine monoclonal antibody to DAF. Molecular analysis has shown that H.A. is homozygous for a single nucleotide substitution, C1579→A, at the position 24 bp upstream of the 3′-end of exon 2 of the DAF gene. This substitution causes the activation of a novel cryptic splice site and results in the production of mRNA with a 26 bp deletion. The deletion introduces a reading frame shift and creates a stop codon immediately downstream of the deletion. Translation of mRNA would be terminated at the first amino acid residue of the second short consensus repeat (SCR2) domain (exon 3) of DAF. The functional domains of DAF's complement regulatory activity and the carboxy-terminal signal domains for glycosylphosphatidylinositol (GPI) anchoring are predicted to be lacking in H.A. Thus, there would be no DAF present on the cell surface.

Sign in / Sign up

Export Citation Format

Share Document