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 Previous Article | Table of Contents | Next Article 
Blood, Vol. 94 No. 8 (October 15), 1999:
pp. 2890-2894
The Molecular Basis of a Case of  -Glutamylcysteine Synthetase
Deficiency
By
Ernest Beutler,
Terri Gelbart,
Takahito Kondo, and
Alison T. Matsunaga
From the Department of Molecular and Experimental Medicine, The
Scripps Research Institute, La Jolla, CA; the Department of
Biochemistry and Molecular Biology in Disease, Atomic Bomb Disease
Institute, Nagasaki University School of Medicine, Nagasaki, Japan; and
the Children's Hospital Medical Center, Oakland, CA.
 |
ABSTRACT |
 -Glutamylcysteine synthetase catalyzes the first step in
glutathione synthesis. The enzyme consists of 2 subunits, heavy and
light, with the heavy subunit serving as the catalytic subunit. A
patient with hemolytic anemia and low red blood cell glutathione levels
was found to have a deficiency of -glutamylcysteine synthetase activity. Examination of cDNA from the patient and her mother showed
that she was homozygous and that her mother was heterozygous for a
A T transversion at nt1109 producing a deduced amino acid change of His370Leu. The partial genomic structure of the catalytic subunit of -glutamylcysteine synthetase (GLCLC) was
determined, providing some intron/exon boundaries to make it possible
to sequence an affected part of the coding region from genomic DNA. The
1109AT mutation was not present in the DNA of 38 normal
subjects. In the course of these studies we found a diallelic
polymorphism in nt +206 of an intron and another polymorphism that
consisted of a duplication of a CAGC at cDNA nt1972-1975 in the
3' untranslated region. The 2 polymorphisms were found to be only
in partial linkage disequilibrium.
© 1999 by The American Society of Hematology.
| |
INTRODUCTION |
REDUCED GLUTATHIONE (GSH) is the major
sulfhydryl compound of erythrocytes. It is transported out of
cells1 and resynthesized, giving a turnover half-time of
approximately 3 days.2 Abnormally low levels of this
tripeptide are encountered in glucose-6-phosphate dehydrogenase
deficiency3 and in deficiencies of the 2 enzymes of GSH
synthesis, -glutamylcysteine synthetase4 (-GCS) and
glutathione synthetase.5 Each of these states is associated
with hemolytic anemia.
Numerous cases of glutathione synthetase deficiency have been
documented, and the molecular lesion in some cases has been identified.6,7 In contrast, -GCS deficiency is rare.
Only 5 patients from 4 unrelated families have been documented
previously,4,8-10 and the mutation has not been reported in
any of these cases.
-Glutamylcysteine synthetase consists of 2 subunits, heavy and
light, with the heavy subunit serving as the catalytic subunit, designated -GCSh, and the light subunit, designated
-GCSl, with regulatory functions. These subunits are
encoded by 2 genes, GLCLC and GLCLR. We now report a
sixth patient with this enzyme deficiency and show that the patient is
homozygous for a mutation at cDNA nt 1109 in GLCLC, in which we
identify an AT transversion that predicts a HisLeu
substitution at amino acid 370 of the catalytic subunit. Previously, a
polymorphic trinucleotide repeat was identified in the 5'
untranslated region (UTR).11 We now demonstrate 2 additional polymorphisms, a tetranucleotide insertion in the 3' untranslated region of the gene and a new diallelic intronic polymorphism.
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MATERIALS AND METHODS |
Enzyme Assays
Assays of red blood cell enzymes and intermediates were performed
essentially as described previously,12,13 but
spectrophotometric assays of glycolytic enzymes were performed in an
automated instrument (Thermomax; Molecular Devices, Sunnyvale, CA) at
30°C. RNA and DNA were prepared from peripheral blood leukocytes by
standard methods.
Sequence Analysis
cDNA was prepared from total RNA extracted from a lymphocyte and
monocyte fraction obtained after hypaque/ficoll sedimentation. Five
micrograms of total RNA was incubated with 300 ng of random primers in
a 38 µL system for 10 minutes at 65°C and allowed to cool slowly
at room temperature. First-strand cDNA was synthesized in a buffer
containing 50 mmol/L Tris-HCl, pH 8.3, 75 mmol/L KCl, 3 mmol/L
MgCl2, 1 mmol/L dithiothreitol, 4 mmol/L dNTPs, 0.8 U/µL RNase Block Inhibitor, and 1 U/µL Moloney murine leukemia virus (MMLV)-reverse transcriptase to which the RNA mixture was
added and incubated at 37°C for 1 hour in a final volume of 50 µL. The reaction was terminated with a 5 minute incubation at
90°C. Subsequent polymerase chain reaction (PCR) reactions
contained 2 µL of the reverse-transcribed cDNA.
The human gene encoding the catalytic subunit of -glutamylcysteine
synthetase (GLCLC) was amplified by PCR using the primers from
the GenBank sequence accession no. M90656. Primers are shown in
Table 1. For primers in the 5'
flanking region of the GLCLC gene containing the putative
antioxidant responsive element and the CAAT and TATA signals, accession
no. L39773 was used. All numbering was based on the adenine in the
initiator ATG being designated 1. The cDNA was amplified in 4 fragments, nt  77 to 684, 535 to 1183, 1028 to 1570, and 1527 to
2044. Genomic DNA was used to amplify the 5' flanking region.
Nested primers were used for sequencing. The amplified PCR products
were sequenced on an Applied Biosystems Inc (Foster City, CA) automatic
sequencer using the dideoxy termination method. Some attempts at
amplifying genomic DNA with the cDNA primers yielded products that were
also sequenced and yielded some information on the genomic structure of
the gene.
The 1109AT mutation creates an Alu I restriction
site, and this was used to confirm the mutation. A genomic fragment 282 bp in length was amplified using a sense intronic primer and an antisense cDNA primer shown in Table 2. The
normal and mutant patterns are also described in Table 2. Thirty-eight
control DNA samples from white subjects were examined for the presence of the mutation.
Population Studies
The 3' tetranucleotide insertion polymorphism was studied in 28 subjects with European ancestry. A PCR fragment from nt 1896 to 2044 was amplified (Table 1) from genomic DNA. Amplified products were
either 149 bp without the insertion or 153 bp if the additional 4 bp
were inserted, indicating that no intron was present. On electrophoresis, the presence of heteroduplexes could be seen in those
samples heterozygous for the repeat. Sequencing of the PCR product
confirmed presence or absence of the repeat.
Immunologic Studies
Preparation of recombinant fusion proteins.
Preparation of constructs, purification, and immunization by
recombinant fusion proteins were performed as described.14 Briefly, a 764-bp DNA fragment (865-1628 bp) of full-length -GCS heavy subunit (-GCSh) cDNA15 was obtained by
digestion with Pst I. Once a 12-bp sequence of BamHI
linker had been added to both sides of the 764-bp -GCSh
cDNA, the cDNA was digested with BamHI and the DNA fragment was
subcloned into the corresponding sites of the pGEX-2T vector (Amersham
Pharmacia, Uppsala, Sweden) for expression as a fusion protein of GST.
A 601-bp DNA fragment (nt 181-781) containing the complete coding
sequence of the human -GCS light subunit (-GCSl) was
generated by reverse transcriptase-PCR (RT-PCR) from total
RNA of human glioblastoma T98G cells serving as a template for the
-GCSl cDNA.16 The forward
(5'-CTCGGATCCGAGGAGCTTCGAGACTGTATCC-3') and reverse (5'-GTACCCGGGCCTGGGCTTCTTCAATGTCAGGGAT-3') primers were
designed to facilitate in-frame insertion into the pGEX-2T vector. The human -GCSl-coding sequence was amplified and digested
with BamHI and Sma I and ligated into the
BamHI/Sma I site of pGEX-2T. Recombinant clones were
analyzed by DNA sequencing to verify that the coding sequence of
-GCS was intact and in-frame with GST.17
The plasmid construct containing GST-human -GCS-subunits was
transfected into Escherichia coli JM109 cells. Production of GST-human -GCS-subunit fusion proteins was induced with isopropyl thiogalactoside, and the fusion protein was batch-purified using glutathione-Sepharose 4B beads (Amersham Pharmacia). Polyclonal antibodies against GST-human -GCSh and GST-human
-GCSl were prepared by immunization of these purified
GST-fusion proteins in rabbits with complete Freund adjuvant twice
monthly. The antibodies were specific to -GCSh or
-GCSl and did not show any cross-reactivity.
Western blots.
Packed cells (0.3 mL) were lysed in a final volume of 1 mL of distilled
water. The solution was applied to a diethyl aminoethyl (DEAE)-cellulose membrane and the membrane was washed with
ice-cold 3 mmol/L sodium phosphate buffer, pH 7.0, until there was no
absorbance at 280 nm. The membrane was then treated with 3 mL of 50 mmol/L Tris-HCl, pH 7.5, containing 0.5 mmol/L  -mercaptoethanol, 0.5 mmol/L MgCl2 (buffer A), and 0.3 mol/L NaCl. The eluant was
dialyzed against 1 L of buffer A for 4 hours and then with the same
volume of buffer A for 12 hours. The dialyzed sample was centrifuged at
18,500g for 10 minutes at 4°C, and the supernatant was
concentrated to be 0.025 mL using a Speed Vac Concentrator (Savant,
Hicksville, NY). Seventy-five micrograms of the sample was applied to a
sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE; 12.5%), transferred to a nitrocellulose membrane, and immunologically stained using the rabbit polyclonal antibody against GST-human -GCSh and GST-human -GCSl. The bound
antibodies were visualized with an alkaline-phosphatase-coupled second
antibody using a ProtoBlot Kit (Promega, Madison, WI). The protein
concentration was determined according to Redinbaugh and
Turley18 with bovine serum albumin as the standard.
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CASE REPORT |
N.S. is a 14-year-old white female who had a blood count performed for
evaluation of menorrhagia. Her hemoglobin was found to be 12.2 g/dL,
with a mean corpuscular volume (MCV) of 105 µm3. Her mother noted that she becomes jaundiced 2 or 3 times per month. She had been told that she has iron deficiency and had been placed on oral iron supplementation. More extensive investigation was performed, showing the hemoglobin to be 11.9 g/dL, the MCV to be
106 µm3, and the reticulocyte count to be 4.9%.
Platelets were 263,000/µL and white blood cells were 7,800/µL. Her
serum folate level was 10.8 ng/mL, the B12 level was 508 pg/mL, and the
thyroid stimulating hormone (TSH) was 2.12 µIU/mL. The direct Coombs
test was negative, haptoglobin was less than 8 mg/dL, and total
bilirubin was 1.5 mg/dL.
In the newborn period, exchange transfusion was required with a total
bilirubin having increased to 23 mg/dL. During childhood, there had
been 2 episodes of head trauma, 1 of which had been followed by a
seizure. However, results of a neurologic evaluation (including
computerized tomography of the head), an electroencephalogram, and a
sleep study were normal. The patient was considered to have a learning
disability with dyslexia. She has difficulty in concentration, impaired
memory, and an auditory processing delay.
The patient's mother is of Pennsylvania Dutch/German/Swedish descent,
while the father is of Pennsylvania Dutch/German/Swedish/Native American descent. The parents are believed to be half-siblings.
On physical examination, including neurologic appraisal, no
abnormalities were found.
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RESULTS |
Erythrocyte Enzymes and Intermediate Values
The levels of erythrocyte enzymes and GSH are summarized in
Table 3.
cDNA Sequence
cDNA from the patient and her mother were sequenced from 58 to
2044 and only the 1109AT mutation and the polymorphic
insertion were found to differ from the published sequence. The patient was homozygous for 1109AT and her mother was a heterozygote
for this transversion. The deduced amino acid change is His370Leu. Genomic DNA from 76 control alleles was tested by restriction analysis,
and none contained this mutation. The patient's brother was
heterozygous for the 1109AT mutation.
A Polymorphism in the 3' UTR
A 3' UTR polymorphism, a duplication of the CAGC at cDNA
nt1972-1975, 61 bp 3' to the TAG stop codon, was found in the
patient and her mother. Both were homozygous for this insertion. The
DNA of 26 normal white subjects was examined for the presence of the CAGC insertion (Table 4).
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Table 4.
Distribution of CAGC Insertion Genotypes and the
Diallelic Polymorphism in the 3' UTR Among 26 Normal White Subjects
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Genomic Sequence and Structure
Genomic DNA from the patient and her mother were sequenced in the
region of the putative antioxidant responsive element and the CAAT and
TATA regions of the GLCLC promoter. No abnormalities were noted.
We have not determined the entire genomic structure of GLCLC,
but in the course of defining the mutation in this patient, 3 introns
were sequenced in their entirety. These sequences have been deposited
in GenBank (accession no. AF118846), and the sequences flanking exons
are shown in Fig 1. An intronic
polymorphism was discovered at position +206 of the last intron shown
in Fig 1. The propositus, her mother, and brother all had the G/G
genotype. The DNA of 26 white subjects was examined for this
polymorphism as well as for the insertional polymorphisms in the
3' UTR (see above). The results of this analysis are shown in
Table 4. The gene frequency of the G allele at nt +206 was 0.79; that
of the A allele was 0.21. The insert in the 3' UTR had a gene
frequency of 0.62; the absence of the insert, then, had a gene
frequency of 0.38. The G allele was significantly associated with the
presence of the insert in the 3' UTR ( 2 = 15; DF = 4; P = .004). However, as shown in Table 4, the intronic polymorphism was not in complete linkage disequilibrium with the 3' UTR polymorphism. Crossovers had clearly occurred.
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| Fig 1.
Partial sequences of three GLCLC introns. The
complete 2,076-bp sequence has been deposited in GenBank (Accession no.
AF118846). cDNA is indicated by upper case letters and introns are
shown as lower case letters.
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Comparison With Other Species
The amino acid sequence around the histidine 370 mutated in our patient
was compared with the published sequences of mouse (Genbank accession
no. P97494) and rat (P19468). Human, mouse, and rat had identical amino
acid sequences in the entire region from amino acid 361 through 475. The sequence surrounding amino acid 370 is shown in
Fig 2. Using the GAP program
(Wisconsin Package Version 9.1, Genetics Computer Group
[GCG], Madison, WI) with the default settings,
comparisons were also made with all of the more distantly related
species for which a sequence of a homolgous enzyme was known, viz
Schizosaccharomyces pombe (S59234), Leishmania
tarentolae (CAA71144), Trypanosoma brucei (AAC47195), Saccharomyces cerevisiae (S59234), Plasmodium falciparum
(CAA07354), Onchocerca volvulus (AAB96970), and
Caenorhabditis elegans (CAA90955). Astonishingly, in each case,
a histidine was aligned with the histidine in the human sequence.
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| Fig 2.
The human, mouse, and rat amino acid sequence surrounding
the amino acid mutated in the patient, histidine 370. The mutated amino
acid is shown in bold type. The 3 sequences are identical in this
region and extending all the way to amino acid 475.
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DISCUSSION |
The synthesis of GSH in red blood cells occurs in 2 steps. First, in a
reaction catalyzed by -glutamylcysteine synthetase, a peptide bond
is formed between the -carboxyl group of glutamic acid and the amino
group of cysteine, forming -glutamylcysteine. Next, glutathione
synthetase catalyzes the formation of a peptide bond between the
carboxyl group of the cysteine of -glutamylcysteine and glycine. In
each of these reactions, a molecule of ATP is dephosphorylated to ADP
and the rate of reaction can conveniently be measured by estimating the
release of inorganic phosphate from --labeled ATP.13
Genetically determined deficiencies of both enzymes of glutathione
synthesis are known. Glutathione synthetase deficiency results in
hemolytic anemia and, in some cases in 5-oxoprolinuria, with associated
neurologic deficits.6 -Glutamylcysteine synthetase deficiency also is known to cause hemolytic anemia. The first reported
patient also had spinocerebellar degeneration,4,19 and for
a time it was believed that this was a part of the syndrome. The fact
that subsequent cases, including the 1 reported here, have been free of
these neurological findings casts doubt on this relationship. However,
it must be admitted that, in the case of a number of genetic disorders
of the red blood cell, variants with and without neurological stigmata
occur, particularly in glutathione synthetase deficiency20
and NADH-diaphorase deficiency.21 The patient reported here
was somewhat mentally retarded, but the relationship to the enzyme
deficiency is not clear. She had neonatal hyperbilirubinemia and
repeated head trauma, and was the offspring of a consanguineous
marriage, so that other autosomal recessive disorders might be expected
to be present.
-Glutamylcysteine synthetase consists of 2 subunits, a heavy
catalytic subunit and a light, regulatory subunit.11,22,23 Its complete sequence has not been determined, and we document here the
sequence of several introns, making it possible to amplify the portion
of the gene in which this mutation occurred, enabling us to use genomic
DNA to screen other samples. Several polymorphisms have been detected
in the GLCLC gene. In addition to a previously documented
trinucleotide repeat in the 5' UTR,11 we found a tetranucleotide repeat in the 3' UTR sequence of the gene and a
diallelic substitution in an intron. The latter 2 polymorphisms were in
partial linkage disequilibrium. The fact that some crossovers had
occurred suggests that these closely spaced polymorphisms are quite
ancient or that crossing over in this region occurs with an unusually
high frequency.
In the patient reported here, we sequenced the entire coding region of
the catalytic subunit and found her to be homozygous for a mutation
1109AT (H370L). The mutation resulted in a marked decrease in
the amount of enzyme protein in the erythrocytes, as indicated by
Western blots (Fig 3). Because we were able to isolate cDNA from a
reticulocyte lysate from the patient and the amount produced was
similar to that from normal lysates, the mutant allele is transcribed.
Presumably, it is also translated, which suggests that the mutant
subunit is unstable. The smaller amount of -GCSl subunit
found on Western blot analysis suggests that the -GCSl
may be stabilized normally by the -GCSh subunit and that, in the absence of the latter, be proteolyzed.
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| Fig 3.
Panel L is stained for the regulatory subunit
(-GCSl), which has a molecular weight of 28,000 (arrow).
Panel H is stained for the catalytic subunit (-GCSh),
which has a molecular weight of 73,000 (arrow). The first lane is a
molecular weight marker: 175,000, 83,000, 62,000, 47,500, 32,500, 25,000, 16,500, and 6,500, respectively, from the top. Lanes 2 and 3, hemolysate from a normal subject; lane 4, hemolysate from the patient;
lanes 5 and 6, hemolysates from the mother and brother of the patient,
respectively. A nonspecific band observed on the third lane of both
sheets may represent an aggregation product formed during preparation
of the samples.
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The parents of the propositus were believed to be half-siblings,
explaining the otherwise unexpected homozygosity for a rare mutant
allele. The sequence abnormalities in the other 4 reported families
have not been reported.
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ACKNOWLEDGMENT |
The authors gratefully acknowledge the assistance of Jennifer
Bojanowski, MS, the genetic counselor, who was of great help in
obtaining the family history and ensuring understanding of the defect;
we acknowledge the cooperation of the family; and we thank Megumi
Sakamoto for her excellent technical assistance.
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FOOTNOTES |
Submitted February 24, 1999; accepted June 15, 1999.
Supported by National Institutes of Health Grants No. HL25552 and
RR00833 and the Stein Endowment Fund. This is manuscript no. 12255-MEM.
The publication costs of this
article were defrayed in part by
page charge payment. This article
must therefore be hereby marked
"advertisement"
in accordance with 18 U.S.C. section
1734 solely to indicate this fact.
Address reprint requests to Ernest Beutler, MD, Department of Molecular
and Experimental Medicine, The Scripps Research Institute, 10550 N
Torrey Pines Rd, La Jolla, CA 92037.
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REFERENCES |
1.
Lunn G, Dale GL, Beutler E:
Transport accounts for glutathione turnover in human erythrocytes.
Blood
54:238, 1979[Abstract/Free Full Text]
2.
Dimant E, Landberg E, London IM:
The metabolic behavior of reduced glutathione in human and avian erythrocytes.
J Biol Chem
213:769, 1955[Free Full Text]
3.
Beutler E, Dern RJ, Flanagan CL, Alving AS:
The hemolytic effect of primaquine. VII. Biochemical studies of drug-sensitive erythrocytes.
J Lab Clin Med
45:286, 1955
4.
Konrad PN, Richards FI, Valentine WN, Paglia DE:
Gamma-glutamyl-cysteine synthetase deficiency.
N Engl J Med
286:557, 1972
5.
Boivin P, Galand C, Andre R, Debray J:
Anémies hémolytiques congenitales avec déficit isole en glutathion réduit par déficit en glutathion synthétase.
Nouv Rev Fr Hematol
6:859, 1966
6.
Dahl N, Pigg M, Ristoff E, Gali R, Carlsson B, Mannervik B, Larsson A, Board P:
Missense mutations in the human glutathione synthetase gene result in severe metabolic acidosis, 5-oxoprolinuria, hemolytic anemia and neurological dysfunction.
Hum Mol Genet
6:1147, 1997[Abstract/Free Full Text]
7.
Shi ZZ, Habib GM, Rhead WJ, Gahl WA, He XW, Sazer S, Lieberman MW:
Mutations in the glutathione synthetase gene cause 5-oxoprolinuria.
Nat Genet
14:361, 1996[Medline]
[Order article via Infotrieve]
8.
Hirono A, Iyori H, Sekine I, Ueyama J, Chiba H, Kanno H, Fujii H, Miwa S:
Three cases of hereditary nonspherocytic hemolytic anemia associated with red blood cell glutathione deficiency.
Blood
87:2071, 1996[Abstract/Free Full Text]
9.
Beutler E, Moroose R, Kramer L, Gelbart T, Forman L:
Gamma-glutamylcysteine synthetase deficiency and hemolytic anemia.
Blood
75:271, 1990[Abstract/Free Full Text]
10.
Richards FI, Cooper MR, Pearce LA, Cowan RJ, Spurr CL:
Familial spinocerebellar degeneration, hemolytic anemia, and glutathione deficiency.
Arch Intern Med
134:534, 1974[Abstract/Free Full Text]
11.
Walsh AC, Li W, Rosen DR, Lawrence DA:
Genetic mapping of GLCLC, the human gene encoding the catalytic subunit of gamma-glutamyl-cysteine synthetase, to chromosome band 6p12 and characterization of a polymorphic trinucleotide repeat within its 5' untranslated region.
Cytogenet Cell Genet
75:14, 1996[Medline]
[Order article via Infotrieve]
12.
Beutler E:
Red Cell Metabolism: A Manual of Biochemical Methods. New York, NY, Grune & Stratton, 1984
13.
Beutler E, Gelbart T:
Improved assay of the enzymes of glutathione synthesis: Gamma-glutamylcysteine synthetase and glutathione synthetase.
Clin Chim Acta
158:115, 1986[Medline]
[Order article via Infotrieve]
14.
Iida T, Mori E, Mori K, Goto S, Urata Y, Oka M, Kohno S, Kondo T:
Co-expression of gamma-glutamylcysteine synthetase sub-units in response to cisplatin and doxorubicin in human cancer cells.
Int J Cancer
682:405, 1999
15.
Kondo T, Yoshida K, Urata Y, Goto S, Gasa S, Taniguchi N:
t-Glutamylcysteine synthetase and active transport of glutathione S-conjugate are responsive to heat shock in K562 erythroid cells.
J Biol Chem
268:20366, 1993[Abstract/Free Full Text]
16.
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
185:29, 1992[Medline]
[Order article via Infotrieve]
17.
Schick C, Kamachi Y, Bartuski AJ, Cataltepe S, Schechter NM, Pemberton PA, Silverman GA:
Squamous cell carcinoma antigen 2 is a novel serpin that inhibits the chymotrypsin-like proteinases cathepsin G and mast cell chymase.
J Biol Chem
272:1849, 1997[Abstract/Free Full Text]
18.
Redinbaugh MG, Turley RB:
Adaptation of the bicinchoninic acid protein assay for use with microtiter plates and sucrose gradient fractions.
Anal Biochem
153:267, 1986[Medline]
[Order article via Infotrieve]
19.
Smith JE, Lee MS, Mia AS:
Decreased gamma-glutamylcysteine synthetase: The probable cause of glutathione deficiency in sheep erythrocytes.
J Lab Clin Med
82:713, 1973[Medline]
[Order article via Infotrieve]
20.
Beutler E:
Glutathione deficiency, pyroglutamic acidemia and amino acid transport.
N Engl J Med
295:441, 1976[Medline]
[Order article via Infotrieve]
21.
Vieira LM, Kaplan J-C, Kahn A, Leroux A:
Four new mutations in the NADH-cytochrome b5 reductase gene from patients with recessive congenital methemoglobinemia type II.
Blood
85:2254, 1995[Abstract/Free Full Text]
22.
Misra I, Griffith OW:
Expression and purification of human gamma-glutamylcysteine synthetase.
Prot Express Purif
13:268, 1998[Medline]
[Order article via Infotrieve]
23.
Mulcahy RT, Wartman MA, Bailey HH, Gipp JJ:
Constitutive and beta-naphthoflavone-induced expression of the human gamma-glutamylcysteine synthetase heavy subunit gene is regulated by a distal antioxidant response element/TRE sequence.
J Biol Chem
272:7445, 1997[Abstract/Free Full Text]
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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]
[PDF]
|
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|
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|
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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.
[Abstract]
[Full Text]
[PDF]
|
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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.
[Abstract]
[Full Text]
[PDF]
|
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|
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E. Ristoff, C. Augustson, J. Geissler, T. de Rijk, K. Carlsson, J.-L. Luo, K. Andersson, R. S. Weening, R. van Zwieten, A. Larsson, et al.
A missense mutation in the heavy subunit of gamma -glutamylcysteine synthetase gene causes hemolytic anemia
Blood,
April 1, 2000;
95(7):
2193 - 2196.
[Abstract]
[Full Text]
[PDF]
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TOP
ABSTRACT
INTRODUCTION