From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
Neonatal Hemolytic Anemia Due to Inherited Harderoporphyria:
Clinical Characteristics and Molecular Basis
By J. Lamoril, H. Puy, L. Gouya, R. Rosipal, V. Da Silva, B. Grandchamp, T. Foint, B. Bader-Meunier,
J.P. Dommergues, J.C. Deybach, and Y. Nordmann
Porphyrias, a group of inborn errors of heme synthesis, are
classified as hepatic or erythropoietic according to clinical
data and the main site of expression of the specific enzymatic defect. Hereditary coproporphyria (HC) is an acute
hepatic porphyria with autosomal dominant inheritance
caused by deficient activity of coproporphyrinogen III oxidase (COX). Typical clinical manifestations of the disease are
acute attacks of neurological dysfunction; skin photosensitivity may also be present. We report a variant form of HC
characterized by a unifying syndrome in which hematologic
disorders predominate: harderoporphyria. Harderoporphyric
patients exhibit jaundice, severe chronic hemolytic anemia
of early onset associated with hepatosplenomegaly, and
skin photosensitivity. Neither abdominal pain nor neuropsychiatric symptoms are observed. COX activity is markedly
decreased. In a first harderoporphyric family, with three
affected siblings, a homozygous K404E mutation has been
previously characterized. In the present study, molecular
investigations in a second family with neonatal hemolytic
anemia and harderoporphyria revealed two heterozygous
point mutations in the COX gene. One allele bore the
missense mutation K404E previously described. The second
allele bore an A=G transition at the third position of the
donor splice site in intron 6. This new COX gene mutation
resulted in exon 6 skipping and the absence of functional
protein production. In contrast with other COX gene defects
that produce the classical hepatic porphyria presentation,
our data suggest that the K404E substitution (either in the
homozygous or compound heterozygous state associated
with a mutation leading to the absence of functional mRNA
or protein) is responsible for the specific hematologic clinical
manifestations of harderoporphyria.
r 1998 by The American Society of Hematology.
H
new case of harderoporphyria bringing new insights into the
clinical and molecular basis of the disease.
UMAN PORPHYRIAS ARE a group of inborn errors of
heme biosynthesis that are classified as hepatic or
erythropoietic according to clinical data and the main site of
expression of the specific enzymatic defect.1 Hereditary coproporphyria (HC) is an autosomal dominant acute hepatic porphyria with incomplete penetrance due to a partial deficiency of
coproporphyrinogen III oxidase (COX; EC 1.3.3.3). COX is a
mitochondrial enzyme2,3 that catalyzes the sixth step in heme
biosynthesis, the decarboxylation of coproporphyrinogen III to
protoporphyrinogen IX.4 Typical clinical manifestations of the
disease resemble two other forms of inherited acute hepatic
porphyria, acute intermittent porphyria (AIP), and variegate
porphyria (VP).5 These porphyrias are characterized by acute
attacks of neurologic dysfunction with abdominal pain, hypertension, tachycardia, and peripheral neuropathy. Skin photosensitivity may also be present in HC and VP. Excretion of large
amounts of coproporphyrin III, mostly in feces and in urine, is
observed.1 COX activity is decreased to 50% of normal controls
in all tissues from coproporphyric patients as well as from
asymptomatic carriers of the gene defect.1 Human cDNA
encoding COX has been sequenced,6,7 and the COX gene
structure has been determined.8,9 To date, eight different mutations have been characterized, which are distributed all over the
COX gene. These mutations, either in the heterozygous (n 5 7)
or homozygous state (n 5 1), are responsible for typical
HC.3,9-13
Harderoporphyria is an erythropoietic variant form of HC
that is biochemically characterized by marked overproduction
in the erythrocytes and increased fecal excretion of the tricarboxylic porphyrin called harderoporphyrin and a markedly decreased lymphocyte COX activity. Harderoporphyria was first
diagnosed in three siblings from healthy nonconsanguineous
parents mainly on the basis of neonatal hemolytic anemia and
skin photosensitivity.14 Molecular studies in the family identified a lysine to glutamic acid susbtitution (K404E) produced by
a homozygous A to C transition at position 1210 in exon 6 of the
COX gene.11 In the present study, we describe and investigate a
Blood, Vol 91, No 4 (February 15), 1998: pp 1453-1457
MATERIALS AND METHODS
Case report. The patient was born at term of healthy, nonconsanguineous French parents. Shortly after birth, he developed severe
jaundice. Physical findings included hepatosplenomegaly and hypospadias. The total serum bilirubin level was 243 µmol/L. The hemoglobin
level was 11.9 g/dL. The nucleated cell count was 160 3 109/L, of
which 85% were erythroblasts. After four exchange transfusions,
performed between the 10 and 91 hours of life, partial regression of
hepatosplenomegaly and resolution of icterus were observed. At 3
months of age, the child was investigated. A blood smear showed 14%
erythroblasts and basophilic stippling. Hemoglobin electrophoresis,
erythrocyte enzyme activities, globin chain synthesis, and immulogic
investigations were normal. Bone marrow aspirate was normal. Hepatic
biopsy showed significant iron storage in hepatocytes without any other
abnormality. At 2 and 7 years of age, the patient was again examined.
Erythrocyte thermal sensitivity was normal and osmotic fragility
increased. Spectrin examination findings were normal. Another bone
marrow examination showed hyperplastic marrow with 50% erythro-
From the Centre Français des Porphyries, INSERM U409, Hôpital
Louis Mourier, Colombes, France; the Department of Pediatrics,
Faculty of Medicine I, Charles University, Praha, Czech Republic; the
Service de Dermatologie, Centre Hospitalier, La Flèche, France; and
the Service de Pédiatrie, Hôpital Kremlin Bicêtre, Kremlin Bicêtre,
France.
Submitted May 28, 1997; accepted October 10, 1997.
Supported by grants from INSERM (U409), University Paris VII in
collaboration with Charles University of Praha (Czech Republic), and
Association Française contre les Myopathies.
Address reprint requests to Y. Nordmann, MD, Laboratoire de
Biochimie, Hôpital Louis Mourier, 92701 Colombes Cedex, France.
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.
r 1998 by The American Society of Hematology.
0006-4971/98/9104-0011$3.00/0
1453
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
1454
LAMORIL ET AL
blasts without dyserythropoiesis. Perls’ Prussian blue stain showed 46%
sideroblasts without ring sideroblasts. Ultrastructural bone marrow
morphology was normal. The hematologic data are summarized in
Table 1. Growth and development remained normal despite persistent
hemolytic anemia and mild splenomegaly. At 8 years of age, skin
fragility, thickening, and erosion of the back of both hands appeared
intermittently without evidence of precipitating factors.
Hepatic porphyria was suspected only when the patient was 18 years
old because of skin lesions associated with chronic hemolytic anemia.
The cutaneous lesions, characterized by the formation of vesicles and
bullae up to 2 cm in diameter, which crusted over and took several
weeks to heal, were localized on light-exposed areas of the backs of the
hands and on the arms and face. The patient had increased skin fragility,
but no hypertrichosis, alopecia, or porphyrin-rich gall stones were found.
The patient is the second of two siblings (Fig 1). In both parents and
his sister, hematologic data were normal (Table 1). The proband and his
relatives never exhibited abdominal and/or neurologic symptoms typical of acute hepatic porphyrias.
Porphyrin synthesis investigations (Table 2). Erythrocyte, urinary,
and fecal porphyrins were determined using standard methods.15,16
Lymphocyte COX activity was measured as described.17
DNA preparation and amplification by polymerase chain reaction
(PCR). Genomic DNA from the proband, his parents, and his sister
was extracted from peripheral blood according to a standard protocol.18
Genomic DNA fragments of interest were amplified by PCR using
primers selected from the published COX sequence.9,12 Twenty picomoles of each set of primers was mixed in 50 µL of PCR solution
containing 1 U of Taq polymerase (Beckmann Inc, Fullerton, CA), 50
mmol/L KCl, 10 mmol/L Tris-HCl, pH 8.5, 1.5 mmol/L MgCl2, and 200
mmol/L of each dNTP. Reactions were performed in a DNA thermocycler (Hybaid, Teddington, UK) as follows: 35 cycles of denaturation at
94°C for 30 seconds, annealing at specific temperature for 30 seconds,
and elongation at 72°C for 1 minute.
Reverse transcription PCR (RT-PCR). Total RNA was extracted
from isolated peripheral blood mononuclear cells using a standard
technique.19 cDNA was obtained by reverse transcription of total RNA
Table 1. Hematologic Data From the Proband and His Family
Age
BasoReticuloErythro
philic
cyte
blastosis MCV
Hb
(%)
(fL) Count (%) Stippling
(g/dL)
Patient Birth 11.2*
3 mo 6†
85
14
64
2 yr
9
50
6 yr
9.2
59
285,000
8%
169,000
5%
25,000
,0.6%
37,000
,0.6%
32,000
,0.6%
19 yr
11
60
50 yr
14.2
85
Mother 49 yr
13.9
80
Sister
13.8
78
Father
84,000
7%
500,000
15%
22 yr
Bone
Marrow
Examination
1
Hyperplasia
Sideroblasts‡:
46%
Hyperplasia
Sideroblasts‡:
30%
Abbreviations: Hb, hemoglobin; MCV, mean corpuscular volume.
*Exchange transfusions were performed.
†A transfusion was performed.
‡No ring sideroblasts.
Fig 1. Family pedigree (solid symbols, patient). In parenthesis is
the lymphocyte COX activity expressed as picomoles of protoporphyrin per hour per milligram of protein at 37°C (normal control value,
350 6 80; mean 6 2 SD).
using oligo(dT) as a primer. cDNA was amplified as already described.10,12
DNA sequencing. All exons and exon/intron boundaries of the
COX gene were amplified using previously selected primers.9,12 PCR
products were purified with the Wizard PCR preps DNA purification
system (Promega-Biotech, Madison, WI). Genomic DNA and cDNA
fragments were directly sequenced using 35S-dATP and the fMol DNA
sequencing kit (Promega-Biotech).
Construction and prokaryotic expression of normal and mutated
human COX cDNA. Normal human cDNA was expressed using the
pGEX-2T expression vector (Pharmacia LKB Biotechnology Inc,
Uppsala, Sweden) as already described.11 To study mutated cDNA with
the exon 6 deletion, site-directed mutagenesis was performed using
normal cloned COX cDNA (pGEX-2T:COX) as template. We used the
Transformer site-directed mutagenesis kit (Clontech Laboratories, Palo
Alto, CA), which is based on the long primer-unique site elimination
mutagenesis method described by Deng and Nickoloff.20 Briefly, long
primers were generated by PCR. 58-Phosphorylated sense oligonucleotide (mutagenic primer), which bypasses exon 6 (105 bp), has 22-bp and
20-bp matching sequences, respectively, flanking the 58 and 38 sides of
the deleted exon. An antisense oligonucleotide which mutates a single
BsaAI restriction site in the pGEX-2T plasmid (selection primer) was
used. The sequences of these primers are as follows: mutagenic primer
(Del.exon6), 58GGCAGCAGCT CAGAAGAGGACG< ATGGGAGTACATGCATTCAC (the arrow indicates the exon 6 bypass); and selection
primer, 58ACACTCCGCTATCGCTCCGCGACTGGGTCATGGCT
(mutated bases abolishing the single BsaAI restriction site are in bold
and underlined).
Standard DNA elongation, ligation, and two-step digestion/transformation of mutated plasmids in mutS Escherichia coli and E coli DH5a
strains were performed according to the manufacturer’s recommendations. The entire sequence of the mutated plasmid was verified by
sequencing. The recombinant bacteria (E coli DH5a) were grown and
COX activities in bacteria lysates were determined as previously
described.21
RESULTS
The patient displayed symptoms and signs of severe hemolytic anemia with splenomegaly and compensatory hyperactive
bone marrow features. In this proband, an atypical profile of
porphyrin excretion was found in feces with massive accumula-
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
HEMOLYTIC ANEMIA AND INHERITED HARDEROPORPHYRIA
1455
Table 2. Porphyrin Concentration in Urine, Feces, and Erythrocytes of Patient and His Parents
Erythrocytes
Subject
Father
Mother
Patient
Normal controls
Urine
Feces
Age
(yr)
Proto
(nmol/L)
ALA
(µmol/L)
PBG
(µmol/L)
URO
(nmol/L)
Copro
(nmol/L)
Total Porphyrins
(nmol)*
Copro
(%)
Hardero
(%)
Proto
(%)
50
49
19
425
402
3,340
,1,900
27
38
40
,38
8
8
9
,5
39
35
120
,50
395
247
1,820
,200
202
104
1,159
,200
17
22
6.2
,22
ND
ND
90
ND
83
71
3
,75
Abbreviations: ALA, aminolevulinic acid; PBG, porphobilinogen; Copro, coproporphyrin; Hardero, harderoporphryin; URO, uroporphyrin;
Proto, protoporphyrin; ND, not detected.
*Results are expressed per gram dry weight.
tion of harderoporphyrin (Table 2). COX activity compared
with control values was decreased by 78% in the patient’s
lymphocytes and by 50% and 30% in those of the father and
mother, respectively (Fig 1).
Sequencing of the seven exons and intron-exon boundaries of
the COX gene from the patient showed two mutations in the
heterozygous state, each on a different allele. The numbering of
the mutations is based on the first base of the initiation codon
described by Delfau-Larue et al.9
The first mutation, an A to G transition at nucleotide 1210,
had been identified in the first reported harderoporphyric
patients.11 This mutation resulted in a lysine to glutamate
substitution at position 404 in the abnormal protein (K404E).
The second mutation was found to be an A to G transition at the
third position of the donor splice site in intron 6 (127713A=G).
This mutation is responsible for exon 6 skipping. After amplification of cDNA from the proband, PCR products showed two
bands, one of the expected size and the other corresponding to a
105-bp deletion in accordance with exon 6 skipping (Fig 2).
Sequencing of the cDNA confirmed the exon 6 deletion that
corresponds to an in-frame deletion of 35 amino acids in the
abnormal protein. Procaryotic expression studies of the exon-6–
deleted cDNA are summarized in Table 3. The enzymatic
activity of the K404E mutated COX protein had already been
investigated.11 In the proband, no other abnormality was found
in the coding sequence.
Direct sequencing of exon 6 and its intron junctions from the
proband’s relatives’ genomic DNAs showed that the father was
heterozygous for the splice site mutation (127713A=G),
whereas the mother and the sister were heterozygous for the
K404E missense mutation.
porphyrias, have not been seen in harderoporphyric patients. In
both harderoporphyric families, the parents were clinically
asymptomatic but exhibited slightly abnormal fecal porphyrin
excretion and an approximately 50% reduction in COX lymphocyte activity.
In the first harderoporphryic cases, molecular studies showed
a homozygous point mutation (A to G transition at nucleotide
1210 in exon 6 of the COX gene) resulting in a lysine to
glutamic acid substitution (K404E).11 The mutated K404E
protein expressed in a procaryotic system showed abnormal
kinetics with reduced affinity, less stability, and a decreased
DISCUSSION
In this study, we report clinical and molecular investigations
in a second family with harderoporphyria. The proband had an
early onset porphyria with severe neonatal hemolytic anemia.
The pattern of porphyrin excretion showed that the major part of
fecal porphyrin was harderoporphyrin, while a large amount of
coproporphyrin was found in urine; in addition, protoporphyrin
was increased in erythrocytes. Enzymatic studies of COX
activity in lymphocytes showed a markedly decreased activity
compatible with a homozygous deficient COX gene. Harderoporphyric patients reported to date (this case and Nordmann et
al14) exhibited strictly identical clinical symptoms characterized
by early onset of hemolytic anemia associated with chronic
cutaneous manifestations. It must be emphasized that abdominal pain and neurologic symptoms, suggestive of acute hepatic
Fig 2. Analysis of RT-PCR products from lymphocyte mRNAs.
Amplified fragments encompassing exon 5, exon 6, and the coding
part of exon 7 were obtained from lymphocyte cDNAs by RT-PCR
using primers HUCO-2-Bio-A and HUCO-10S (9) and analyzed on 2%
agarose gel. Two amplified products were obtained from the heterozygous harderoporphyric patient (P), the 468-bp fragment containing
the K404E missense mutation and the 363-bp fragment resulting from
the mRNA with complete deletion of exon 6. Amplification of control
mRNA (N) showed only the normal 468-bp fragment. M, molecular
size markers.
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
1456
LAMORIL ET AL
Table 3. Expression of the Exon-6–Deleted COX cDNA in E coli
Normal control
Negative control
Exon-6–deleted cDNA
Construct
COX Activity*
pGEX-2T:COX/sense
pGEX-2T:COX/antisense
pGEX-2T:COX/del. exon 6
15,870
1,317
629
Expression vector: pGEX-2T. Values are expressed as the mean of
two duplicate experiments.
*Activity given as picomoles of protoporphyrin per hour per milligram at 37°C.
residual activity (25% of control). It has been suggested that the
two decarboxylation steps catalyzed by COX take place at the
same catalytic site and that this mutation was localized at this
active site of the enzyme.11 Consistent with this hypothesis,
COX from harderoporphyric patients has been found to have a
similarly increased Km for both coproporphyrinogen and
harderoporphyrinogen substrates.14
In this study, we show that harderoporphyria, occurring in a
second family, resulted from compound heterozygous mutations affecting the COX gene. The previously reported K404E
mutation was found in the heterozygous state in the proband, his
mother, and his sister. The second mutation, an exon 6 skipping
mutation caused by an A to G transition at position 127713, is a
new mutation that resulted in an in-frame deletion of 35
aminoacids in the mutated protein. This mutation was found in
the heterozygous state in the proband and his father. The father
did not exhibit acute porphyria syndrome typical of HC,
probably because of the low penetrance of the disease, especially in males. Expression studies showed that the truncated
protein encoded by the exon-6–deleted COX mRNA had
virtually no residual enzymatic activity (Table 3). Interestingly,
a previous exon 6 skipping mutation (not assessed with
expression studies) has been reported in a patient of Czech
origin with a heterozygous G to A transition at position 1277,
the last position of the splice donor site of exon 6.9 This patient
had a typical clinical form of HC. He repeatedly exhibited
neurologic symptoms with paresis, and the diagnosis was made
during hospitalization after treatment with barbiturates.9 All
these data indicate that the residual enzyme activity found in
compound heterozygous harderoporphyric patients results exclusively from the K404E mutated COX protein. Therefore, the
specific harderoporphyric symptoms appear directly related to
this point mutation in the COX gene.
To date, eight different mutations in the COX gene have been
characterized. They were responsible for typical HC.3 Only one
homozygous form of HC has been reported,10 but its clinical
and biologic presentation was completely different from harderoporphyria. The patient had a clinical history of severe acute
attacks of hepatic porphyria, without chronic hemolytic anemia,
a large accumulation in feces of coproporphyrin with harderoporphyrin being absent, and a profound defect of COX activity
in lymphocytes.22 Molecular investigations showed an arginine
to tryptophan substitution (R231W) in exon 5 of the COX gene.10
It has been hypothesized that the active COX protein acts as a
homodimer of approximatively 70 to 74 kD.23,24 Because of the
lack of crystallographic data, little structural information about
the human COX enzyme is available. Recently, a histidine
residue at position 258 has been shown to be a highly conserved
region of aerobic COX; it could be involved in COX catalytic
activity through a hypothetic and controversial interaction with
Cu21.25,26 Our studies on harderoporphyria show that the lysine
residue at position 404 is also important for catalytic activity of
the enzyme: the K404E mutation is probably responsible for
accumulation of harderoporphyrinogen, an intermediate in the
oxidative decarboxylation of coproporphyrinogen. It has been
suggested that this intermediate would leave the abnormal
enzyme more easily and, after spontaneous oxidation to harderoporphyrin, would accumulate in the patient.11 Moreover, comparison of nucleotide deduced amino acid sequences from
humans, Saccharomyces cerevisiae, Salmonella typhimurium, E
coli,26-29 and mouse30 showed that the K404E mutation occurred
in a region highly conserved throughout evolution (Table 4). Our
data and the high percentage of conserved aminoacids suggest
that exon 6 may play an important role in the catalytic activity
and/or maintenance of the active conformation of the enzyme.
The pathogenesis of the hematologic symptoms in harderoporphyria is not yet fully understood. However, as observed in
erythropoietic porphyrias (Günther’s disease, erythro-hepatic
porphyria), harderoporphyric patients exhibit splenomegaly.
The spleen is the major site for removal of damaged or
hemolyzed erythrocytes31; hence, the splenomegaly observed in
harderoporphyria could be presumed to be secondary to this
process. Extrinsic abnormalities of erythrocytes seem unlikely,
because the direct Coomb’s test was negative and the survival
time of normal erythrocytes transfused into harderoporphyric
patients was normal. The overproduction of porphyrins in
harderoporphyria may account for the hemolytic symptoms.
The elevated level of erythrocyte protoporphyrin found in all
the harderoporphyric patients, in contrast with classical HC,
provides evidence favoring the bone marrow as a source of
Table 4. Comparison of Amino Acid Sequences Deduced From Nucleotides Sequences of the Human (HC), From Codon 387 to 448,
Mouse (MC), Saccharomyces cerevisiae (SC), E coli (EC), Salmonella typhimurium (ST), and Soybean (GM)
HC
MC
SC
EC
ST
GM
LRRG
LRRG
IRRG
YRRG
YRRG
LRRG
***
K404E
<
RYVEFNLLYDRGTKFGLFTPGSRIESILMSLPLT
RYVEFNLLYDRGTKFGLFTPGSRIESILMSLPLT
RYVEFNLIYDRGTQFGLRTPGSRVESILMSLPEH
RYVEFNLVWDRGTLFGLQT-GGRTESILMSMPPL
RYVEFNLVWDRGTLFGLQT-GGRTESILMSMPPL
RYVEFNLVYDRGTTFGLKT-GGRIESILVSLPLT
******* **** *** * * * **** * *
ARWEYMHSPSENSKEAEILEVLRHPRDWV
ARWEYMHSPSENSKEAEILEVLRHPRDWV
ASWLYNHHPAPGSREAKLLEVTTKPREWV
VRWEYDYQPKDGSPEAALSE-FIKVRDWV
VRWEYDWQPEAGSPEAALSE-FIQVRDWI
ARWEYDHKPEEGSEEWKLLDACINPKEWI
***
*
* *
*
Deleted aminoacid sequence deduced from exon 6 deletion nucleotides sequence is boxed. Mutated amino acid (K404) encoded by A1210G
allelic mutation is represented above the human sequence. Asterisks indicate identical amino acids.
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
HEMOLYTIC ANEMIA AND INHERITED HARDEROPORPHYRIA
elevated porphyrins in this disease and could be involved, at
least in part, in the hemolytic process. Hemolysis of erythrocytes may also result from photolysis as porphyrin-laden cells
are exposed to light in the dermal capillaries. Light wavenlengths suitable for porphyrin photoactivation are known to
penetrate the skin to a depth sufficient to produce this phenomenon and photohemolysis has also been demonstrated in vitro.32,33
In conclusion, this study suggests for the first time the
existence of a phenotype/genotype relationship in the human
COX gene. In contrast with other COX gene defects responsible
for HC, the K404E mutation in the homozygous state or
associated with a deleterious allele (exon 6 skipping in this
case) induces harderoporphyria. The abnormal kinetic pattern
with reduced affinity, less stability, and the decreased (25%)
residual activity of the mutated K404E is responsible for the
unusual accumulation of harderoporphyrin and the specific
hematologic and clinical symptoms of harderoporphyria. Harderoporphyria is a unifying syndrome of childhood onset with
clinical features quite different from those observed in other
hepatic porphyrias. It is characterized by jaundice, hemolytic
anemia, hepatosplenomegaly, skin photosensitivity, and a marked
increase in harderoporphyrin in urine and feces.
REFERENCES
1. Kappas A, Sassa S, Galbraith RA, Nordmann Y: The porphyrias,
in Scriver CR, Beaudet AL, Sly WS, Valle D (eds): The Metabolic and
Molecular Basis of Inherited Disease. New York, NY, McGraw-Hill,
1995, p 2103
2. Grandchamp B, Phung N, Nordmann Y: The mitochondrial
localization of coproporphyrinogen III oxidase. Biochem J 176:97, 1978
3. Grandchamp B, Lamoril J, Puy H: Molecular abnormalities of
coproporphyrinogen oxidase in patient with hereditary coproporphyria.
J Bioenerg Biomembr 27:215, 1995
4. Elder GH, Evans JO, Jackson JR, Jackson AH: Factors determining the sequence of oxidative decarboxylation of the 2-and 4-propionate
substituents of coproporphyrinogen III by coproporphyrinogen III
oxidase in rat liver. Biochem J 169:215, 1978
5. Nordmann Y, Deybach JC: Human hereditary porphyrias, in
Dailey, HA (ed): Biosynthesis of Heme and Chlorophylls. New York,
NY, McGraw-Hill, 1990, p 491
6. Martasek P, Camadro JM, Delfau-Larue MH, Dumas JB, Montagne JJ, De Verneuil H, Labbe P, Grandchamp B: Molecular cloning
sequencing and functional expression of a cDNA encoding human
coproporphyrinogen oxidase. Proc Natl Acad Sci USA 91:3024, 1994
7. Taketani S, Kohno H, Furukawa T, Yoshinaga T, Tokunaga R:
Molecular cloning, sequencing and expression of cDNA encoding
human coproporphyrinogen oxidase. Biochem Biophys Acta 1183:547,
1994
8. Cacheux V, Martasek P, Fougerousse F, Delfau MH, Druart L,
Tachdjian G, Grandchamp B: Localization of the coproporphyrinogen
oxidase gene to chromosome band 3q12. Hum Genet 94:557, 1994
9. Delfau-Larue MH, Martasek P, Grandchamp B: Coproporphyrinogen oxidase: Gene organization and description of a mutation leading to
exon skipping. Hum Mol Genet 3:1325, 1994
10. Martasek P, Nordmann Y, Grandchamp B: Homozygous hereditary coproporphyria caused by an arginine to tryptophan substitution in
coproporphyrinogen oxidase and common intragenic polymorphisms.
Hum Mol Genet 3:477, 1994
11. Lamoril J, Martasek P, Deybach JC, Da Silva V, Grandchamp B,
Nordmann Y: A molecular defect in coproporphyrinogen oxidase gene
causing harderoporphyria, a variant form of hereditary coproporphyria.
Hum Mol Genet 4:275, 1995
12. Lamoril J, Deybach JC, Puy H, Grandchamp B, Nordmann Y:
1457
Three novel mutations in the coproporphyrinogen oxidase gene. Hum
Mutat 9:78, 1997
13. Fugita H, Kondo M, Taketani S, Nomura N, Furuyama K, Akagi
R, Nagai T, Terajima M, Galbraith RA, Sassa S: Characterization and
expression of cDNA encoding coproporphyrinogen oxidase from a
patient with hereditary coproporphyria. Hum Mol Genet 3:1807, 1994
14. Nordmann Y, Grandchamp B, De Verneuil H, Phung L: Harderoporphyria: A variant hereditary coproporphyria. J Clin Invest 72:1139,
1983
15. Bissel DM: Laboratory evaluation in porphyria. Semin Liver Dis
2:100, 1982
16. Lim CK, Li F, Peters TJ: High performance liquid chromatography of uroporphyrinogen and coproporphyrinogen isomers with amperometric detection. Biochem J 234:629, 1986
17. Guo R, Lim CK, Peters TJ: Accurate and specific HPLC assay of
coproporphyrinogen III activity in human peripheral leucocytes. Clin
Chim Acta 177:245, 1988
18. Loparev VN, Cartas MA, Monken CE, Velpandi A, Srinivasan A:
An efficient and simple method of DNA extraction from whole blood
and cell lines to identify infectious agents. J Virol Methods 34:105, 1991
19. Chomczynski P, Sacchi N: Single step method of RNA isolation
by acid guanidium thiocyanate-phenol-chloroform extraction. Anal
Biochem 162:156, 1987
20. Deng WP, Nickoloff JA: Site-directed mutagenesis of virtually
any plasmid by eliminating a unique site. Anal Biochem 200:81, 1992
21. Grandchamp B, Nordmann Y: Decreased lymphocyte coprorporphyrinogen III oxidase activity in hereditary coproporphyria. Biochem
Biophys Res Commun 74:1089, 1978
22. Grandchamp B, Phung N, Nordmann Y: Homozygous case of
hereditary coproporphyria. Lancet 2:1348, 1977
23. Bogard M, Camadro JM, Nordmann Y, Labbe P: Purification and
properties of mouse liver coproporphyrinogen oxidase. Eur J Biochem
181:417, 1989
24. Camadro JM, Chambon H, Jolles J, Labbe P: Purification and
properties of coproporphyrinogen oxidase from the yeast Saccharomyces cerevisiae. Eur J Biochem 156:579, 1986
25. Kohno H, Furukawa T, Tokunaga R, Taketani S, Yoshinaga T:
Mouse coproporphyrinogen oxidase is a copper-containing enzyme:
Expression in Escherichia coli and site-directed mutagenesis. Biochem
Biophys Acta 1292:156, 1996
26. Medlock AE, Dailey HA: Human coproporphyrinogen oxidase is
not a metalloprotein. J Biol Chem 271:32507, 1996
27. Troup B, Jahn M, Hungerer C, Jahn D: Isolation of the hemF
operon containing the gene for the Escherichia coli aerobic coproporphyrinogen III oxidase by in vivo complementation of a yeast HEM13
mutant. J Bacteriol 176:673, 1994
28. Xu K, Elliott T: An oxygen-dependent coproporphyrinogen
oxidase encoded by the hemF gene of Salmonella typhimurium. J
Bacteriol 175:4990, 1993
29. Zagorec M, Buhler JM, Treich I, Keng T, Guarente L, LabbeBois R: Isolation, sequence, and regulation by oxygen of the yeast
HEM13 gene coding for coproporphyrinogen oxidase. J Biol Chem
263:9718, 1988
30. Kohno H, Furukawa T, Yoshinaga T, Tokunaga R, Taketani S:
Coproporphyrinogen oxidase purification, molecular cloning, and induction of mRNA during erythroid differentiation. J Biol Chem 268:21359,
1993
31. Haining RG, Cowger ML, Shurtleff DB, Labbe RF: Congenital
erythropoietic porphyria. Special studies and therapy. Am J Med
45:624, 1968
32. Nordmann Y, Deybach JC: Congenital erythropoietic porphyria.
Semin Liver Dis 2:154, 1982
33. Sassa S, Kappas A: The porphyrias, in Harris H, Hirschorn K
(eds): Advances in Human Genetics. New York, NY, Plenum, 1981, p
121
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
1998 91: 1453-1457
Neonatal Hemolytic Anemia Due to Inherited Harderoporphyria: Clinical
Characteristics and Molecular Basis
J. Lamoril, H. Puy, L. Gouya, R. Rosipal, V. Da Silva, B. Grandchamp, T. Foint, B. Bader-Meunier, J.P.
Dommergues, J.C. Deybach and Y. Nordmann
Updated information and services can be found at:
http://www.bloodjournal.org/content/91/4/1453.full.html
Articles on similar topics can be found in the following Blood collections
Red Cells (1159 articles)
Information about reproducing this article in parts or in its entirety may be found online at:
http://www.bloodjournal.org/site/misc/rights.xhtml#repub_requests
Information about ordering reprints may be found online at:
http://www.bloodjournal.org/site/misc/rights.xhtml#reprints
Information about subscriptions and ASH membership may be found online at:
http://www.bloodjournal.org/site/subscriptions/index.xhtml
Blood (print ISSN 0006-4971, online ISSN 1528-0020), is published weekly by the American Society of
Hematology, 2021 L St, NW, Suite 900, Washington DC 20036.
Copyright 2011 by The American Society of Hematology; all rights reserved.