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Next Article 
Blood, Vol. 95 No. 7 (April 1), 2000:
pp. 2193-2196
PLENARY PAPER
A missense mutation in the heavy subunit of  -glutamylcysteine
synthetase gene causes hemolytic anemia
Ellinor Ristoff,
Camilla Augustson,
Judy Geissler,
Thea de Rijk,
Katarina Carlsson,
Jia-Li Luo,
Kerstin Andersson,
Ron S. Weening,
Rob van Zwieten,
Agne Larsson, and
Dirk Roos
From the Department of Pediatrics, Karolinska Institute, Huddinge
University Hospital, Huddinge, Sweden; the Central Laboratory of the
Netherlands Red Cross Blood Transfusion Service and Laboratory of
Experimental and Clinical Immunology, Academic Medical Center,
University of Amsterdam, Amsterdam, The Netherlands; the Department of
Anesthesiology, Karolinska Institute, Huddinge University Hospital,
Huddinge, Sweden; and the Emma Children's Hospital, Academic Medical
Center, University of Amsterdam, Amsterdam, The Netherlands.
 |
Abstract |
 -Glutamylcysteine synthetase (GCS) catalyzes the initial and
rate-limiting step in the biosynthesis of glutathione. -GCS consists
of a heavy and a light subunit encoded by separate genes. Hereditary
deficiency of GCS has been reported in 6 patients with hemolytic anemia
and low erythrocyte levels of glutathione and -glutamylcysteine. In
addition, 2 patients also had generalized aminoaciduria and developed
neurologic symptoms. We have examined a Dutch kindred with 1 suspected
case of GCS deficiency. The proband was a 68-year-old woman with a
history of transient jaundice and compensated hemolytic anemia. One of
her grandchildren was also GCS deficient; he was 11 years old and had a
history of neonatal jaundice. The enzyme defect was confirmed and GCS
activity was found to be less than 2% of normal in the erythrocytes of
both patients. The complementary DNA (cDNA) for the heavy subunit of GCS was sequenced in these patients and in several members of the
family. The proband and her GCS- deficient grandson were identified as homozygous for a 473C T substitution, changing codon 158 from CCC for proline into CTC for leucine. Several family members with half-normal GCS activity in their erythrocytes were heterozygous for
the mutation.
(Blood. 2000;95:2193-2196)
© 2000 by The American Society of Hematology.
| |
Introduction |
Glutathione (-glutamyl-cysteinyl-glycine) (GSH) is
present in all mammalian tissues. It is found intracellularly in
millimolar concentrations, in tissues such as the liver, kidney, brain,
skeletal muscle, and erythrocytes. Glutathione plays a key role in many biologic functions, such as protein and DNA synthesis, detoxification of xenobiotics and free radicals, as well as reductive
reactions.1
The turnover occurs via the -glutamyl cycle, which involves 6 enzymes.2 Glutathione is synthesized by the consecutive action of -glutamylcysteine synthetase (glutamate-cysteine ligase) (GCS) and glutathione synthetase (GS). The rate-limiting step in the
biosynthesis is catalyzed by GCS, which is feedback-inhibited by GSH.
This enzyme is a dimer consisting of a heavy (GCSh) and a
light (GCSl) subunit. The human gene for GCSh,
called GLCLC, has been localized to chromosome 6p123 and
the gene for GCSl, called GLCLR, to chromosome
1p21.4 The heavy subunit (molecular weight, 72.6 kd)
exhibits the catalytic activity of the native enzyme and is also
responsible for the feedback inhibition by GSH. The light subunit
(molecular weight, 27.7 kd) is catalytically inactive, but plays an
important regulatory role by lowering the Km of GCS for
glutamate and raising the Ki for GSH.5,6
Hereditary GCS deficiency has been reported in 6 patients.7-11 The sixth patient was recently reported and
in this patient the first molecular diagnosis was made.11
We found 2 additional patients with the same disorder in a large Dutch
family with hereditary GSH deficiency in the erythrocytes, originally
described by Prins et al.12 We identified the molecular
mutation on the heavy subunit causing hereditary GCS deficiency in this family.
| |
Materials and methods |
This study was approved by the Ethics Committee of Karolinska Institute.
Patient history
Patient 1:2.
The first patient was originally reported more than 30 years ago with a
GSH deficiency in her erythrocytes (Figure
1).12 This proband is a Dutch
woman born in 1931. Between the ages of 20 and 30, she had had at least
4 episodes of jaundice, and when investigated in her thirties she had
hemolytic anemia and low erythrocyte GSH levels (2% of the normal
mean). Four other members of her family had transient jaundice and
marked erythrocyte GSH deficiency. One of them also had mild
hepatosplenomegaly.12 Since the age of 30, the proband has
had well-compensated hemolytic anemia and episodes of jaundice when
eating fat food. The jaundice has no known relation to medication or
infections. This patient and her family refrain from eating fava beans.
She has also complained of a vestibular organ defect with equilibrium
dysfunction, which was also present in some of her GSH-deficient
siblings. When investigated at 68 years of age, she had the following
laboratory findings: mild anemia with hemoglobin 6.5 mmol/L (reference
values, 7.4-9.6 mmol/L), erythrocytes 3.4 × 109/L
(reference values, 3.7-5.0 × 109/L); mean
corpuscular volume 94 fL (reference values, 83-105 fL). According to
the family doctor, the patient is mentally normal and has no neurologic
complaints.
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| Fig 1.
Pedigree of the Dutch GCS-deficient family.
Patient 1:2 was previously reported as Mrs. M.-K. (patient V:19) with
GSH deficiency in the erythrocytes (Prins et al12). Her
parents were consanguineous (second cousins). Subjects 2:2 and 2:6 are
sisters, also related to patient 1:2 (their father is a second cousin
of patient 1:2).
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Patient 3:10.
This grandson of patient 1:2 is the first child of consanguineous
parents (Figure 1). He had a history of neonatal jaundice requiring an
exchange transfusion. Because the jaundice was more severe than normal,
he was subjected to a liver biopsy, which had normal histology. During
childhood, he had a well-compensated hemolytic anemia, but no other symptoms.
Analyses of -glutamyl cycle enzymes
Preparation of hemolysates.
Packed erythrocytes were obtained from EDTA blood samples by
centrifugation at 900g for 3 minutes after washing 3 times with 5 volumes of cold isotonic NaCl solution. The packed erythrocytes were
lysed by the addition of 1 volume of 50 mmol Tris-HCl buffer (pH
7.4)/L, containing 1 mmol EDTA/L, and by sonication for 2 × 20
seconds. The erythrocyte membranes were removed by centrifugation at
18,000g for 40 minutes. Hemoglobin was determined using
Vanzetti's method.13
Preparation of leukocyte extracts.
Leukocyte extracts were obtained from EDTA blood samples by lysis of
erythrocytes in a lysis buffer containing 155 mmol ammonium chloride/L,
10 mmol sodium bicarbonate/L, and 0.1 mmol EDTA/L. The EDTA blood
sample was mixed with 40 mL cold lysis buffer and put on ice for 15 minutes. The erythrocytes were removed by centrifugation at
800g for 5 minutes. The leukocyte pellet was resuspended in 5 mL cold lysis buffer and put on ice for 10 minutes. The cell suspension
was diluted to 50 mL with cold isotonic saline solution and centrifuged
at 800g for 5 minutes. The pellet was again washed with
isotonic saline and centrifuged at 1600g for 10 minutes. The
pellet with leukocytes was analyzed.
Assay of enzyme activities and thiol-bimane adducts.
Activity of -GCS and levels of glutathione and -glutamylcysteine
were assayed by the method described by Luo et al14 with the following modifications. The incubation mixture (final volume 300 µL) contained 100 mmol Tris-HCl (pH 8.2)/L, 6 mmol adenosine triphosphate (ATP)/L, 50 mmol KCl/L, 6 mmol dithiothreitol (DTT)/L, 20 mmol MgCl2/L, 3 mmol cysteine/L, and 15 mmol glutamic
acid/L. The mixture was preincubated at 37°C for 15 minutes to
ensure the complete reduction of disulfides to thiols. The reaction was initiated by the addition of various amounts of cell-free extracts from
erythrocytes or leukocytes. After 20 minutes of incubation at 37°C,
the reaction was stopped by adding 10 µL of 80% 5-sulfosalicylic acid (SSA). After removal of denaturated protein by centrifugation at
12,000g for 15 minutes, samples of the supernatant solution (100 µL) were derivatized to detect the reaction product,
-glutamylcysteine. For the blank sample, 10 µL SSA was added to
the incubation buffer before adding hemolysate or leukocyte lysate,
respectively. The samples to be derivatized were added to 100 µL of
20 mmol monobromobimane/L (in 50 mmol N-ethylmorpholine/L). The mixture
was placed in the dark at room temperature for 10 minutes. The reaction
was stopped by adding 10 µL of 80% SSA. The high-performance liquid
chromatography separation of the thiol-bimane adducts on a
reversed-phase Supercosil LC-18 octadecylsilyl silica column was
followed by fluorimetric detection, as reported
elsewhere.15
Glutathione synthetase was determined by the method described by
Wellner et al16 with the following modifications. The
incubation volume was 100 µL and the mixture contained 12 mmol
L--glutamyl-l- -aminobutyrate/L, 16 mmol
1-14C-glycine/L (specific activity, 9.25 MBq/mmol)
including 0.4 µCy/L, 4 mmol phosphoenolpyruvate/L, 4 mmol sodium
ATP/L, 8000 U pyruvate kinase/L, 100 mmol Tris HCl-buffer (pH 8.6)/L,
25 mmol KCl/L, 6 mmol MgCl2/L, and 1 g bovine serum
albumin/L. The reaction was started by the addition of different
amounts of cell extracts. After incubation at 37°C for 120 minutes,
the reaction was stopped by the addition of 10 µL of 20% perchloric
acid. Denatured protein was removed by centrifugation at
18,000g for 2 minutes, and the supernatant solution was
transferred to a 0.5 (diameter) × 4 (height) cm Dowex
1 × 8 acetate ion exchange column. The remaining
1-14C-glycine was eluted with 6 mL of 20 mmol HAc/L and
14C-ophthalmic acid (the product) was eluted with 6 mL of
1.5 mol ammonium acetate/L. The radioactivity was analyzed in a liquid scintillation counter.
Mutation analysis.
RNA was isolated from leukocytes by the method of Chirgwin et
al.17 cDNA was synthesized, as described
previously.18 The coding region of the GCSh
gene (GLCLC) was amplified by polymerase chain reaction (PCR) in 7 overlapping fragments with a set of oligonucleotide
primers.19 Each sense-forward primer contained a
"flag," that is, a 20-nucleotide  21M13 phage sequence
(5'-TGTAAAACGACGGCCAGT-3') for BIGDYE primer
cycle sequencing (ABI Prism, Applied Biosystems, Perkin-Elmer
Biosystems, Foster City, CT). The positions of the primers
complementary to the GLCLC cDNA sequence were:
F1 sense (4-23) 5'-flag-ACGAGGCTGAGTGTCCGTC-3'
F1 antisense (369-388) 5'-CTCCCATACTCTGGTCTCCA-3'
F2 sense (293-312) 5'-flag-GGTCCTGTCTGGGGAGAAAG-3'
F2 antisense (638-657) 5'-AGGTACTGAAGCGAGGGTGC-3'
F3 sense (581-600) 5'-flag-CCCAGTGGAAGGAGGAGCT-3'
F3 antisense (950-969) 5'-TGTCTGACACATAGCCTCGG-3'
F4 sense (923-942) 5'-flag-GGCTTTGAGTGCTGCATCTC-3'
F4 antisense (1321-1340) 5'-TGGAGGAGGGGGCTTAAATC-3'
F5 sense (1191-1210) 5'-flag-CTGGCCCAGCATGTTGCTCA-3'
F5 antisense (1572-1591) 5'-CCATCCACCACTGCATTGCC-3'
F6 sense (1519-1538) 5'-flag-GAGATGCTGTCTTGCAGGGA-3'
F6 antisense (1931-1950) 5'-CAAGTAACTCTGGGCATTCA-3'
F7 sense (1680-1699) 5'-flag-GTGTTTCCTGGACTGATCCC-3'
F7 antisense (2114-2133) 5'-ATTTCTGGCTCACTGGCCCA-3'
The PCR was performed in an Air Thermo Cycler (type 1605, Idaho
Technology, Idaho Falls, ID) with cDNA as a template, 50 nmol/L (for
fragment 1, 60 nmol/L) of each primer, 0.8 mmol/L of each dNTP, 1 U of
Gold Taq DNA polymerase (HT Technologies, Amsterdam, The Netherlands),
and 90 nmol/L antiTaq (Clontech Laboratories, Palo Alto, CA) in a
buffer recommended by the manufacturer (HT Technologies), in a total
volume of 15 µL in glass capillaries. Each PCR cycle consisted of a
10-second denaturation at 95°C, 45-second annealing at 55°C
(fragments 2 and 3 at 60°C), and 30-second elongation at 72°C.
In total, 50 cycles were run. Nucleotide sequences of the PCR products
were determined directly by BigDye Primer Cycle Sequencing (Applied
Biosystems) in an ABI Prism 377 sequencer (Perkin Elmer Biosystems).
Polymorphisms in codon 158 were excluded by fragment analysis after
digestion of PCR product 2 with the restriction enzyme Bsa JI (New
England Biolabs, Beverly, MA). The wild-type PCR product with the CCC
codon 158 was digested into 3 fragments of 274, 92, and 16 nucleotides;
the PCR product with the CTC codon 158 was digested into 2 fragments of
274 and 108 nucleotides. The fragments were separated by agarose-gel
electrophoresis and detected under UV light.
| |
Results |
Patient 1:2 had less than 2% of the normal GCS activity in her
erythrocytes (Figure 1 and Table 1). The GS
activity was normal. The erythrocyte GS also had normal affinity for
its substrates -glutamylcysteine and glycine (not shown). The
erythrocyte GSH and -glutamylcysteine level was less than 1% of
normal (Table 1). In the leukocytes of patient 1:2, the GCS activity
was less than 1% of normal and the GSH level was 7% of normal.
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Table 1.
Content of -glutamyl cycle enzymes and products
in erythrocytes and leukocytes of 2 GCS-deficient patients
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The GCS activity in hemolysates from patient 1:2 did not increase
markedly with an increase in glutamate concentration (data not shown).
Patient 1:2 had normal levels of amino acids in her blood and urine and
did not excrete excessive amounts of 5-oxoproline in her urine.
Patient 3:10 was found to have a GCS activity of less than 1% of
normal in his erythrocytes (Figure 1 and Table 1). The GS activity was
normal. The erythrocyte glutathione level was 1.5% of normal,
and the erythrocyte -glutamylcysteine was less than 1% of normal.
Both patients were found to be homozygous for a 473CT
mutation in GLCLC, predicting a Pro158Leu substitution in
this protein (Figure 2). This mutation was
not present in 70 GCSh alleles from control subjects. The
GCS activity in erythrocytes was distinctly different in family members
with no mutation (23.9,22.7-27.5 pkatal/mg Hb [median, range];
n = 6) and family members who were heterozygous carriers
(13.1,12.4-14.9; n = 8) or homozygous patients (< 0.1 and 0.412;
n = 2) (Figure 1). The GSH content of erythrocytes in family members
who were heterozygous carriers and members with no mutation was normal,
as was the GS activity.
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| Fig 2.
Analysis of cDNA from patient 3:10, his parents, and his
siblings.
A PCR product containing the region around 473C of the GLCLC gene was
generated from cDNA and was analyzed by Dye Primer Cycle sequencing.
The results show (arrows) that the parents 2:5 and 2:6 are both
heterozygous for the 473CT substitution in the GLCLC gene,
patient 3:10 is homozygous for this mutation, his sister 3:11 is
normal, and his brother 3:12 is heterozygous.
|
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| |
Discussion |
We studied a large inbred family and found 2 people with GCS deficiency
and hemolytic anemia. Other symptoms may have been coincidental.
We obtained indirect evidence that the hereditary defect was located in
the heavy, catalytic subunit of GCS and not in the light, regulatory
subunit, because the activity of the mutant GCS could not be normalized
by raising the glutamate concentrations markedly6 (data not
shown). Therefore, we started our molecular analysis by sequencing the
cDNA of the heavy subunit. In both patients we found a homozygous
473CT mutation in the GLCLC gene, predicting a
Pro158Leu substitution in the GCSh subunit.
Genetic analysis also revealed that several other family members were heterozygous for this mutation. All of these heterozygotes had erythrocyte GCS activity reduced to about half-normal. Therefore, it is
very likely that the Pro158Leu substitution in the
GCSh subunit is the cause of the decreased GCS activity.
The heterozygous family members had neither decreased levels of GSH nor
any clinical symptoms. For this reason we conclude that a mutation of
the GLCLC gene in a heterozygous state does not cause hemolytic anemia
or other symptoms.
Six patients have hitherto been reported with hereditary GCS deficiency
and all show similar symptoms (Table 2). The first 2 were siblings of German descent, a brother and a sister, who in their
late twenties developed hemolytic anemia, markedly decreased survival
time of erythrocytes, erythrocyte GSH deficiency, and generalized
aminoaciduria (patients 1 and 2 in Table 2).7,8 The sister
became psychotic after treatment with sulfonamide for a urinary tract
infection. In their thirties, both developed spinocerebellar degeneration and a neuromuscular disorder with peripheral neuropathy and myopathy. The sister required blood transfusions during pregnancy, but otherwise the increased rate of hemolysis was well compensated in
both patients. Both showed normal activity of GS.
The third patient having GCS deficiency was a woman of Polish
extraction with consanguineous parents (patient 3 in Table
2).9 She had a history of transient jaundice complicating
an unspecified viral infection at 10 years of age. At 21, she developed
anemia and reticulocytosis while pregnant and required blood
transfusions. After the delivery, she was still anemic, with high
reticulocyte counts. She was found to have an erythrocyte GSH
deficiency and markedly diminished GCS and GSH-S-transferase activity.
She had normal activity of GS. No neurologic abnormality was detected. The GSH-S-transferase deficiency has been postulated to be secondary to
the lack of GSH.20
Two unrelated Japanese patients with GCS deficiency have been described
(patients 4 and 5 in Table 2).10 A 15-year-old boy had a
history of anemia at birth and had received a blood transfusion at 1 month of age. Thereafter he had been asymptomatic until 10 years of
age, when he had an episode of anemia, reticulocytosis, jaundice, and
hepatosplenomegaly. At 15 years of age, he still had mild hemolytic
anemia. His growth and development were normal. He showed no signs of
central nervous system involvement. He had markedly diminished activity
of GCS and GSH-S-transferase, but normal activity of GS. His mother
also had mild deficiencies of GCS and GSH-S-transferase. The fifth
patient was a 17-year-old girl with consanguineous parents. The girl
had had anemia and reticulocytosis since childhood. She had no
aminoaciduria (J. Ueyama and A. Hirono, personal communication, 1998).
She had a markedly decreased erythrocyte survival time. Her erythrocyte GCS activity was decreased, as also was her GSH-S-transferase activity. The activity of GS was normal. Her mother also had mild deficiencies of GCS and GSH-S-transferase.
Recently (published after submission of our manuscript) a sixth patient
with GCS deficiency was described by Beutler et al (patient 6 in Table
2).11 This patient was a 14-year-old white girl of
Pennsylvania-Dutch/German/Swedish/Native American descent. Her parents
were related to each other. The girl was examined because of
menorrhagia, anemia, and reticulocytosis. She also became jaundiced 2 or 3 times per month. She had a history of neonatal hyperbilirubinemia
requiring exchange transfusion. During childhood she had had 2 episodes
of head trauma, 1 of which had been followed by a seizure. She was
considered to have a learning disability with dyslexia and was also
thought to be mentally retarded. She had, however, no abnormalities by
physical examination, including neurologic evaluation. She was found to
have an erythrocyte GSH deficiency and markedly diminished GCS and
GSH-S-transferase activity, but normal activity of GS. Her mother and
brother had intermediate activity of GCS but normal GSH levels. The
patient was homozygous for an 1109AT mutation, in the GLCLC
gene, predicting a His370Leu substitution in
GCSh. The patient's mother and brother were heterozygous for the same mutation. The level of mutant messenger RNA (mRNA) in the
patient's reticulocytes were normal, whereas the amount of mutant
protein in her erythrocytes was decreased, indicating that the mutant
protein was unstable.
Orlowski and Meister21 have suggested that the -glutamyl
cycle is involved in the active transport of amino acids. -Glutamyl transpeptidase was postulated to play a key role in this process, because it is located on the outer surface of the cell membrane, especially in the epithelia of tissues extensively involved in transport, for example, the kidney. Four of the 8 patients with GCS
deficiency hitherto described have been investigated with respect to
urinary amino acids. Two had generalized aminoaciduria, but 2 did not.
These findings may have several explanations. Assuming that the
-glutamyl cycle is involved in amino acid transport,21 the absence of aminoaciduria could be due to residual low levels of GCS
being sufficient to permit function of the -glutamyl cycle. As with
deficiencies of other erythrocyte enzymes, such as glucose-6-phosphate dehydrogenase22 and cytochrome b5 reductase,23
mutations that cause instability of the enzyme may become manifest
only in the red blood cells, because these cells have a rather long
half-life and lack the capacity to synthesize new protein. In
contrast, mutations that inactivate the catalytic capacity of
GCS may lead to GSH deficiency in all tissues, with severe clinical
consequences. Unfortunately, the 3-dimensional structure of
GCSh is not yet known. It is, therefore, not possible to
speculate about the effect of the proline-158 substitution for leucine
in this protein.
Another explanation for the lack of aminoaciduria in patients with GCS
deficiency may be the existence of alternative pathways for amino acid
transport, not involving the -glutamyl cycle. The aminoaciduria
reported in 2 of 8 patients may then be completely unrelated to the
enzyme defect in the -glutamyl cycle. The present identification of
the gene mutation responsible for GCS deficiency makes it possible to
gain further knowledge about the functions of residual GCS activity in
various tissues. The aminoaciduria may also be secondary to the low
levels of GSH in the cells, similar to the decreased activity of
GSH-S-transferase detected in patients with GS
deficiency.20
From the findings in 8 patients with GCS deficiency, it may be
concluded that all patients have hemolytic anemia and low erythrocyte levels of both GSH and -glutamylcysteine. Our 2 patients with GCS
deficiency were homozygous for a missense mutation in the GCSh subunit. It seems important to determine the
underlying mutations in the other GCS-deficient patients to learn more
about the physiologic properties of GCS. The relationship between
aminoaciduria, neurologic and psychiatric disorders, and the primary
enzyme deficiency remains to be established.
| |
Acknowledgments |
We thank Dr H. A. Wassink for providing us with clinical data,
Associate Professor Jan Wernerman for placing his laboratory facilities
at our disposal, and Mrs Christina Hebert for skillful technical assistance.
| |
Footnotes |
Submitted September 27, 1999; accepted November 30, 1999.
Supported by the Swedish Medical Research Council (project nos. 4792 and 4210), Frimurarebarnhuset Foundation, Wera Ekström Foundation, Samariten Foundation, and Sven Jerring Foundation.
Reprints: Ellinor Ristoff, Department of Pediatrics,
Karolinska Institute, Huddinge University Hospital, S-141 86 Huddinge, Sweden; e-mail: ellinor.ristoff{at}klinvet.ki.se.
The publication costs of this
article were defrayed in part by
page charge payment. Therefore,
and solely to indicate this fact,
this article is hereby marked
"advertisement"
in accordance with 18 U.S.C.
section 1734.
| |
References |
1.
Meister A, Anderson ME.
Glutathione.
Annu Rev Biochem.
1983;52:711[Medline]
[Order article via Infotrieve].
2.
Meister A.
On the enzymology of amino acid transport.
Science.
1973;180:33[Free Full Text].
3.
Sierra-Rivera E, Summar ML, Dasouki M, Krishnamani MR, Phillips JA, Freeman ML.
Assignment of the gene (GLCLC) that encodes the heavy subunit of gamma-glutamylcysteine synthetase to human chromosome 6.
Cytogenet Cell Genet.
1995;70:278[Medline]
[Order article via Infotrieve].
4.
Sierra-Rivera E, Dasouki M, Summar ML, et al.
Assignment of the human gene (GLCLR) that encodes the regulatory subunit of gamma-glutamylcysteine synthetase to chromosome 1p21.
Cytogenet Cell Genet.
1996;72:252[Medline]
[Order article via Infotrieve].
5.
Meister A.
Glutathione metabolism and its selective modification.
J Biol Chem.
1988;263:17,205[Free Full Text].
6.
Huang CS, Chang LS, Anderson ME, Meister A.
Catalytic and regulatory properties of the heavy subunit of rat kidney gamma-glutamylcysteine synthetase.
J Biol Chem.
1993;268:19,675[Abstract/Free Full Text].
7.
Konrad PN, Richards FD, Valentine WN, Paglia DE.
Glutamyl-cysteine synthetase deficiency: a cause of hereditary hemolytic anemia.
N Engl J Med.
1972;286:557.
8.
Richards FD, Cooper MR, Pearce LA, Cowan RJ, Spurr CL.
Familial spinocerebellar degeneration, hemolytic anemia, and glutathione deficiency.
Arch Intern Med.
1974;134:534[Medline]
[Order article via Infotrieve].
9.
Beutler E, Moroose R, Kramer L, Gelbart T, Forman L.
Gamma-glutamylcysteine synthetase deficiency and hemolytic anemia.
Blood.
1990;75:271[Abstract/Free Full Text].
10.
Hirono A, Iyori H, Sekine I, et al.
Three cases of hereditary nonspherocytic hemolytic anemia associated with red blood cell glutathione deficiency.
Blood.
1996;87:2071[Abstract/Free Full Text].
11.
Beutler E, Gelbart T, Kondo T, Matsunaga AT.
The molecular basis of a case of gamma-glutamylcysteine synthetase deficiency.
Blood.
1999;94:2890[Abstract/Free Full Text].
12.
Prins HK, Oort M, Zurcher C, Beckers T.
Congenital nonspherocytic hemolytic anemia, associated with glutathione deficiency of the erythrocytes: hematologic, biochemical and genetic studies.
Blood.
1966;27:145[Abstract/Free Full Text].
13.
Vanzetti G.
An azide-methemoglobin method for hemoglobin determination in blood.
J Lab Clin Med.
1966;67:116[Medline]
[Order article via Infotrieve].
14.
Luo JL, Hammarqvist F, Andersson K, Wernerman J.
Surgical trauma decreases glutathione synthetic capacity in human skeletal muscle tissue.
Am J Physiol.
1998;275:E359[Abstract/Free Full Text].
15.
Luo JL, Hammarqvist F, Cotgreave IA, Lind C, Andersson K, Wernerman J.
Determination of intracellular glutathione in human skeletal muscle by reversed-phase high-performance liquid chromatography.
J Chromatogr B Biomed Appl.
1995;670:29[Medline]
[Order article via Infotrieve].
16.
Wellner VP, Sekura R, Meister A, Larsson A.
Glutathione synthetase deficiency, an inborn error of metabolism involving the gamma-glutamyl cycle in patients with 5-oxoprolinuria (pyroglutamic aciduria).
Proc Natl Acad Sci U S A.
1974;71:2505[Abstract/Free Full Text].
17.
Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ.
Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease.
Biochemistry.
1979;18:5294[Medline]
[Order article via Infotrieve].
18.
Kuijpers TW, Van Lier RA, Hamann D, et al.
Leukocyte adhesion deficiency type 1 (LAD-1)/variant: a novel immunodeficiency syndrome characterized by dysfunctional beta2 integrins.
J Clin Invest.
1997;100:1725[Medline]
[Order article via Infotrieve].
19.
Gipp JJ, Chang C, Mulcahy RT.
Cloning and nucleotide sequence of a full-length cDNA for human liver gamma-glutamylcysteine synthetase.
Biochem Biophys Res Commun.
1992;185:29[Medline]
[Order article via Infotrieve].
20.
Beutler E, Gelbart T, Pegelow C.
Erythrocyte glutathione synthetase deficiency leads not only to glutathione but also to glutathione-S-transferase deficiency.
J Clin Invest.
1986;77:38.
21.
Orlowski M, Meister A.
The gamma-glutamyl cycle: a possible transport system for amino acids.
Proc Natl Acad Sci U S A.
1970;67:1248[Abstract/Free Full Text].
22.
Roos D, van Zwieten R, Wijnen JT, et al.
Molecular basis and enzymatic properties of glucose 6-phosphate dehydrogenase Volendam, leading to chronic nonspherocytic anemia, granulocyte dysfunction, and increased susceptibility to infections.
Blood
1999;94:2955[Abstract/Free Full Text].
23.
Shirabe K, Yubisui T, Borgese N, Tang CY, Hultquist DE, Takeshita M.
Enzymatic instability of NADH-cytochrome b5 reductase as a cause of hereditary methemoglobinemia type I (red cell type).
J Biol Chem.
1992;267:20,416[Abstract/Free Full Text].
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e102 - e105.
[Abstract]
[Full Text]
[PDF]
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I.-S. Song, S. Tatebe, W. Dai, and M. T. Kuo
Delayed Mechanism for Induction of {gamma}-Glutamylcysteine Synthetase Heavy Subunit mRNA Stability by Oxidative Stress Involving p38 Mitogen-activated Protein Kinase Signaling
J. Biol. Chem.,
August 5, 2005;
280(31):
28230 - 28240.
[Abstract]
[Full Text]
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D. Hamilton, J. H. Wu, M. Alaoui-Jamali, and G. Batist
A novel missense mutation in the {gamma}-glutamylcysteine synthetase catalytic subunit gene causes both decreased enzymatic activity and glutathione production
Blood,
July 15, 2003;
102(2):
725 - 730.
[Abstract]
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S.-i. Koide, K. Kugiyama, S. Sugiyama, S.-i. Nakamura, H. Fukushima, O. Honda, M. Yoshimura, and H. Ogawa
Association of polymorphism in glutamate-cysteine ligase catalytic subunit gene with coronary vasomotor dysfunction and myocardial infarction
J. Am. Coll. Cardiol.,
February 19, 2003;
41(4):
539 - 545.
[Abstract]
[Full Text]
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S.-i. Nakamura, K. Kugiyama, S. Sugiyama, S. Miyamoto, S.-i. Koide, H. Fukushima, O. Honda, M. Yoshimura, and H. Ogawa
Polymorphism in the 5'-Flanking Region of Human Glutamate-Cysteine Ligase Modifier Subunit Gene Is Associated With Myocardial Infarction
Circulation,
June 25, 2002;
105(25):
2968 - 2973.
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[Full Text]
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A. C. Walsh, J. A. Feulner, and A. Reilly
Evidence for Functionally Significant Polymorphism of Human Glutamate Cysteine Ligase Catalytic Subunit: Association with Glutathione Levels and Drug Resistance in the National Cancer Institute Tumor Cell Line Panel
Toxicol. Sci.,
June 1, 2001;
61(2):
218 - 223.
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[Full Text]
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Top
Abstract
Introduction