|
|||||||||
|
|
|
|
|
|
|
|
|
|
|
HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
From the Department of Molecular
Patho-Biochemistry and Patho-Biology, Department of Pathology, Yamagata
University School of Medicine, Japan; Department of Internal Medicine
I, Fukushima Prefectural Medical College, Japan; and Department of
Pediatrics, Fukuoka University School of Medicine, Japan.
Two Japanese patients were newly diagnosed as having B subunit
(XIIIB) deficiency of factor XIII (former type I deficiency). Both
patients have a previously described one-base deletion at the boundary
between intron A/exon II in the XIIIB gene, heterozygously or homozygously. A founder effect was proposed for this mutation because 3 unrelated patients with XIIIB deficiency also share 2 3'-polymorphisms. In one patient heterozygous for the above mutation, a
novel mutation was also identified: a deletion of guanosine in exon IX
(delG) of the XIIIB gene. To understand the molecular and
cellular pathology of the delG mutation, expression studies were
performed using a cultured mammalian cell line. Pulse-chase experiments
showed that a resultant truncated XIIIB remained inside the cells and
could not be secreted into the culture medium. Furthermore, immunocytochemical examinations by epifluorescence, confocal, and
electron microscopes indicated impaired intracellular transportation of
the truncated XIIIB from the endoplasmic reticulum to the Golgi apparatus. No mutations in the gene for the A subunit (XIIIA) were
identified in this patient. Therefore, secretion of the truncated XIIIB
must also be impaired in vivo, leading to a secondary XIIIA deficiency.
These results support a previous conclusion that genetic defects of
XIIIB are the basis for the former type I factor XIII deficiency.
(Blood. 2001;97:2667-2672) Factor XIII deficiency is an inherited hemorrhagic
disease characterized by a lifelong bleeding tendency and abnormal
wound healing in affected patients and spontaneous abortion in female patients.1-3 Coagulation factor XIII is a proenzyme of
plasma transglutaminase and circulates in blood as a heterotetramer
consisting of 2 catalytic A subunits (XIIIA) and 2 noncatalytic B
subunits (XIIIB). Factor XIII catalyzes intermolecular cross-linking
reactions between fibrin monomers themselves, fibrin and
More than 200 cases of factor XIII deficiency have been identified. In
most cases, diagnosis of factor XIII deficiency is made by measuring
the enzymatic activity of factor XIII, which represents the amount of
functional XIIIA. Factor XIII deficiency had been classified by the
presence or absence of antigens into 2 categories: type I,
characterized by the lack of both XIIIA and XIIIB; and type II,
characterized by the lack of XIIIA alone.4 Until now, most
cases of factor XIII deficiency were diagnosed as type II and only 3 cases have been reported to be type I.4-7 The reliability
of classification by immunologic assays depends on the specificity of
antibodies against each subunit. From the results of genetic analyses
on factor XIII deficiency, we previously concluded that type I
deficiency results from genetic defects of XIIIB, and type II from
genetic defects of XIIIA. We then proposed a new classification of
factor XIII deficiency at the DNA level: XIIIA deficiency (former type
II deficiency) and XIIIB deficiency (former type I deficiency), and a
possible combined deficiency of XIIIA and XIIIB.8 This
proposal was approved by the Scientific and Standardization Committee
(SSC) in 1999 (http://www.med.unc.edu/isth/99FXIII.html).
We performed genetic analyses in all cases of XIIIB deficiency and
identified 3 mutations in the XIIIB genes. Two mutations were found in one Japanese patient (patient 1): a deletion of adenosine, ag In the present study, 2 Japanese patients were newly diagnosed as
having XIIIB deficiency. Both were found to be heterozygous or
homozygous for the known ag Patients
Venous blood was drawn from normal individuals and patients with XIII
deficiency and their family members after informed consent had been
obtained. Genomic DNA was isolated from peripheral blood cells by
standard techniques.
Immunoblot analysis
In vitro amplification of the XIIIB gene and nucleotide sequence analysis The XIIIB gene of each patient was amplified from 0.5 µg genomic DNA using 3 pairs of specific primers by polymerase chain reaction (PCR) using a LA PCR kit (Takara, Kyoto, Japan). The sequences of specific primers for the XIIIB gene were as follow: for exons I to V (~7 kb), 5'-AAT TCT AGA CTC GTT ACA AAA GGA CTT AGA CAG AGG-3' (sense) and 5'-GAT GGA TCC ACA GGA ATT TTG TCA GAG CTA AT-3' (antisense); for exons VI to X (~7 kb), 5'-TAG CTG CAG TCT GCC ATT CTC TCT ATG ATG-3' (sense) and 5'-CAT AAG CTT AGT GTT ATG TTA ACT CAA AAA TGA CAG C-3' (antisense); for exons XI and XII (~1.7 kb), 5'-CAA GAA TTC CTG TTC TTA CCA AGA TGT AAC AAG -3' (sense) and 5'-TAT GAA TTC TAG TCA ATG GGC ATT AGG AAA TG-3' (antisense). PCR was performed with denaturation at 98°C for 20 seconds, and annealing and extension at 68°C for 10 minutes, for 30 cycles. Nucleotide sequence analysis was carried out with a BigDye Terminator cycle sequencing kit (PE Biosystems, Foster City, CA) using a 310 Genetic Analyzer (PE Biosystems).Screening for mutations in the XIIIB gene by PCR-restriction fragment length polymorphism analysis Three known mutations of the XIIIB gene (ag [-]g,
Cys430-Phe, and AAC insertion) and 2 polymorphisms (one at the
3'-untranslated region [A/G27970] and another in the 3'-flanking
region) were determined as described previously.9,11,12
For detection of a novel mutation (delG), an oligonucleotide with one
base mismatch (in italic) was designed as an antisense primer, 5'-CTC
TGT TGC ACT GCA CAG AGA ATT-3' (E9Gdel). A fragment
containing exon IX was amplified by a combination of E9Gdel and a sense
primer, 5'-ATA AAG CTT TTG AAA CTT GTC ATA ATT TTT GTT TTA-3' (B4-5H).
PCR was carried out with denaturation at 94°C for 30 seconds,
annealing at 55°C for 30 seconds, and extension at 72°C for 30 seconds, 35 times. An amplified product of 260 bp was treated with
EcoRI endonuclease at 37°C for 90 minutes; then the sample
was applied to a 2% agarose gel.
Construction of expression vectors for wild-type and mutant recombinant XIIIBs A full-length complementary DNA (cDNA) of XIIIB was cleaved from an expression vector for wild-type recombinant XIIIB (rXIIIB), ZMB3/rXIIIb,13 and subcloned into the EcoRI site of pBluescript II (Stratagene, La Jolla, CA), to yield pBSII/WT rXIIIB. A region of exon IX containing the novel delG mutation was amplified by PCR using patients' genomic DNA as a template. The amplified fragment was treated with RsaI and BsgI endonucleases, and then a released fragment of 145 bp was inserted into the RsaI-BsgI site of pBSII/WT rXIIIB to yield pBSII/Mut rXIIIB. An EcoRI fragment of the mutant rXIIIB cDNA was then cleaved from the pBSII/Mut rXIIIB and ligated into the EcoRI site of the ZMB3 expression vector to generate ZMB3/Mut rXIIIB.Cell culture and selection of stable transfectants for wild-type and mutant rXIIIBs Baby hamster kidney (BHK) cells were cultured in Dulbecco modified Eagle medium (DMEM; Nikken Biomedical Laboratory, Tokyo, Japan) supplemented with 10% (vol/vol) fetal bovine serum (MBL, Nagoya, Japan), and antibiotics (Gibco BRL, Gaithersburg, MD) at 37°C in a 5% CO2 incubator. Transfection was performed using the modified calcium phosphate precipitation method of Chen-Okayama. Stably transfected cells that expressed the wild-type or mutant rXIIIB were selected as described previously.10Pulse-chase experiments The day before these experiments were performed, rXIIIB-transfected cells (4 × 105 cells) were plated onto a 3-cm dish and incubated in serum-free DMEM without methionine (Met; Sigma, St Louis, MO) for 1 hour at 37°C before radiolabeling. Newly synthesized proteins were labeled with 25 µCi/plate of [35S]-Met in Met-depleted serum-free DMEM for 20 minutes (pulse), and further incubated in serum-free DMEM supplemented with unlabeled Met (chase). Samples of culture media and cell lysates were harvested at varying time intervals and stored immediately at80°C. To remove nonspecificically absorbed proteins,
the samples were pretreated with PANSOLBIN cells (Wako Chemicals,
Tokyo, Japan) alone, and only supernatants were used for
immunoprecipitation. The samples were incubated with a rabbit antihuman
XIIIB antibody for 1 hour at 4°C and incubated further with PANSOLBIN
cells at room temperature for 30 minutes with shaking. The precipitates
were washed 3 × with 2 × Tris-buffered saline (TBS) containing
0.2% Tween 20. Bound antigen was eluted from PANSOLBIN cells by
heating at 100°C for 5 minutes and subjected to 8% SDS-PAGE. The
PAGE gel was dried and exposed to an imaging plate (Fuji Film, Tokyo,
Japan). The exposed imaging plate was analyzed by FLA2000 (Fuji Film)
using MacBAS version 2.x software.
Immunocytochemistry The rXIIIB-transfected cells were washed twice with phosphate-buffered saline (PBS), fixed with 4% paraformaldehyde for 15 minutes, and then permeabilized with 0.1% Triton-X100 for 10 minutes at room temperature. The permeabilized cells were washed twice with PBS, incubated with 2% bovine serum albumin (BSA) in PBS, and reacted with a rabbit antihuman XIIIB antibody at a dilution of 1:1000 in PBS containing 2% BSA for 4 hours at room temperature. The washed cells were incubated for another 2 hours at room temperature with a goat antirabbit IgG conjugated with fluorescein 5-isothiocyanate (FITC; Organon Teknika, Durham, NC) at a dilution of 1:1000 in PBS containing 2% BSA. Immunocytochemical staining was repeated using a mouse monoclonal antibody against Golgi 58K protein (Golgi58; Biomaker, Rehovot, Israel) as a Golgi marker or against protein disulfide isomerase (PDI; Stressgen, Victoria, BC, Canada) as an endoplasmic reticulum (ER) marker, and a rhodamin-conjugated antimouse IgG (ICN, Aurora, OH). The anti-Golgi 58K protein and anti-PDI antibodies were used at dilutions of 1:200 and 1:1000, respectively. After 3 washes with PBS, cover slides were mounted with 0.01% paraphenylendiamine in PBS containing 90% glycerol, and the samples were observed under an epifluorescence or confocal microscope (Olympus IX-70, Tokyo, Japan).Immunoelectron microscopy The rXIIIB-transfected cells were reacted with a rabbit antihuman XIIIB antibody and a goat antirabbit IgG conjugated with FITC by the same procedure as immunocytochemistry, except for the usage of suspended cells. After reacting with an HRP-conjugated rabbit anti-FITC IgG (Dako, Glostrup, Denmark), the cells were washed 3 × with 1% BSA in PBS and incubated with a DAB-hydrogen peroxide solution at room temperature for 10 minutes. After 3 washings with 1% BSA in PBS, the samples were immersed for 10 minutes in 2.5% buffered glutaraldehyde, washed in 0.1% cacodylate buffer (pH 7.4), refixed with 1% osmium tetroxide (pH 7.2) for 1 hour, and washed again in the cacodylate buffer. After dehydration in ethanol, the pieces were embedded in Epon 812 (Nisshin EM, Tokyo, Japan). Ultrathin sections were cut for electron microscopy (EM109, Zeiss, Esslingen, Germany).
Immunoblot analysis of plasma samples To obtain a rough estimate of the amounts of both subunits in each patient, immunoblot analyses of plasma samples were performed using anti-human XIIIA or XIIIB antibodies (Figure 1). XIIIB was scarcely detectable in the patients, although it was clearly detected in a normal sample even at 160 × dilution. On the other hand, XIIIA was detectable in all plasma samples. In patient 4, the XIIIA was significantly decreased to about 12% of normal plasma. The plasma XIIIA level in patient 5 was approximately 40%, although it varied between 5% and 40% through his neonatal period. These results clearly indicate that both patients had complete XIIIB deficiency (former type I factor XIII deficiency).
Search for known mutations in the XIIIB gene We had previously identified 3 mutations (ag()g, Cys430-Phe,
and AAC insertion) in the XIIIB genes of all the known
patients with XIIIB deficiency worldwide (Figure
2, top). To look for the presence of
these mutations, genomic DNAs of these patients were tested by
PCR-restriction fragment length polymorphism (RFLP) analyses. The ag()g mutation, identified in patient 1 heterozygously, creates a recognition site for TaqI
endonuclease,9 so that TaqI-treatment of a
1.8-kb fragment amplified from patient 1 yielded 3 fragments (Figure 2,
bottom): an uncleaved fragment of 1.8 kb for the ag allele, and cleaved
fragments of 1.5 and 0.3 kb for the ()g allele. The same digestion
pattern was observed in patient 4, indicating that patient 4 is a
heterozygote for the ag()g mutation. On the other hand, an
amplified DNA from patient 5 was completely cleaved into 2 fragments of
1.5 and 0.3 kb, indicating that patient 5 was a homozygote for
this mutation.
Neither of 2 other known mutations were detected in patient 4 or 5, although they were observed in the index cases (patients 1 and 2) in whom each mutation had been identified previously (Figure 2, bottom). Genetic relationship among 3 Japanese patients All 3 patients (patients 1, 4, and 5) with the ag()g mutation
are ethnic Japanese but unrelated. The ag()g mutation was not found
in either of 2 Italian patients (patients 2 and 3, data not shown). To
search for the genetic relationship among the 3 Japanese patients, the
ag()g mutation and 2 known polymorphisms in the 3'-region of the
XIIIB gene were determined among the patients and members of
their families (Figure 3). The 2 known
polymorphisms were nucleotide A/G27970 in the 3'-untranslated region,
and a short tandem repeat of (TTTA)n (n = 7-10) in the
3'-flanking region (Figure 2, top). Gene frequencies of these
polymorphisms among 90 normal Japanese individuals were as follow:
G27970 and A27970, 0.813, and 0.187, respectively; and 10-, 9-, and
8-repeats of TTTA, 0.733, 0.217, and 0.050, respectively.
In a homozygote for the ag Identification of a new mutation in patient 4 From the above results, it was predicted that patient 4 had another mutation(s) in the remaining XIIIB allele. Nucleotide sequence analysis of the XIIIB gene was then carried out for patient 4. First, PCR-amplified DNAs were sequenced directly. The ag()g mutation was confirmed to be heterozygous, and
a novel mutation was also found in exon IX. Doublet peaks of the
nucleotide started to appear from the nucleotide position of 14522 in
both sense and antisense directions (data not shown), suggesting the
presence of a deletion/insertion. To confirm this mutation,
PCR-amplified exon IX was subcloned, and 8 subclones were then
sequenced. A novel mutation, the deletion of one guanosine at the
nucleotide position of 14522 (delG), was identified in 3 out of the
8 subclones.
Analysis by PCR-RFLP was also performed to confirm the presence of the
novel delG mutation. A PCR product amplified from the normal allele had
a new EcoRI site, but one from the delG allele did not
(Figure 4A). Therefore, the PCR product
of 260 bp from a normal control would be completely cleaved with
EcoRI endonuclease into 2 fragments of 236 bp and 24 bp,
although the latter would scarcely be detectable because of its small
size. The amplified products from patient 5 and a normal individual
were completely cleaved by the EcoRI treatment (Figure 4B).
However, the PCR product of patient 4 yielded 2 fragments of 236 bp and
24 bp, whereas an uncleaved fragment of 260 bp remained even after
prolonged EcoRI digestion. These results indicate that the
delG mutation was present heterozygously in patient 4. Because the delG
mutation was not found among 90 normal Japanese individuals, it must be a second causative mutation in the remaining XIIIB allele of patient 4, which is responsible for XIIIB deficiency.
Sequencing analyses of all PCR products amplified from patient 5 showed
the homozygous ag Impaired secretion of truncated rXIIIB The delG mutation, newly identified in the XIIIB gene of patient 4, introduces a premature stop codon with a frameshift, resulting in the production of a truncated XIIIB. To understand the molecular and cellular pathology of this novel mutation, we performed expression studies using a mammalian cultured cell line (BHK) and expression vectors for wild-type and delG rXIIIBs. The rXIIIB-expressing cells were selected by expression levels determined by Northern blot and immunoblot analyses (data not shown). In metabolic labeling studies, a wild-type rXIIIB of about 80 kd and a truncated rXIIIB of about 60 kd were detected in the lysates of rXIIIB-transfected cells (Figure 5); thus, the actual size of each band corresponded well to its expected size. No dimer was observed for the mutant rXIIIB under nonreducing conditions, although the eighth Sushi domain of the mutant would contain an odd number of cysteine residues3 and thus one cysteine residue must be left unpaired. These results are in contrast to our previous report in which a Cys430-Phe mutant did form a dimer.10
In pulse-chase experiments, it was shown that the wild-type rXIIIB was
exported from the cells to the medium immediately, and that nearly all
labeled rXIIIB was found in the medium 2 hours after labeling (Figure
6A). The exported rXIIIB was present at a
fairly constant level in the medium for 24 hours (Figure 6B). The
truncated rXIIIB, however, remained inside the cells and could not be
secreted into the medium at all. After 2 hours, the truncated rXIIIB in
the cells was decreased to 50% of that at 0 hour.
Intracellular localization of truncated rXIIIB In general, a secretory protein goes through the ER to the Golgi apparatus before secretion. Therefore, we determined the intracellular localization of the wild-type and mutant rXIIIBs by an immunocytochemical staining method using a rabbit antihuman XIIIB antibody. Stably transfected cells of the wild-type rXIIIB showed a typical staining pattern of secretory proteins (Figure 7A): no signal was detected in mock cells (data not shown). Intense signals for the wild-type rXIIIB were observed mainly near the nuclei, which likely corresponds to the Golgi, and were colocalized with the signals for Golgi 58K protein (Figure 7A, top). In contrast, cells of the mutant rXIIIB were stained in a reticular pattern throughout the cytoplasm (Figure 7A, bottom). The signals for the mutant rXIIIB were colocalized with those for an ER marker protein, PDI. The same patterns of distribution for the wild-type and mutant rXIIIB were also observed by confocal microscopy (data not shown). In addition, immunoelectron microscopic pictures clearly showed accumulation of rXIIIB in the Golgi of BHK cells expressing the wild-type (Figure 7B, top), whereas rXIIIB locates in the rough ER of cells expressing the mutant protein (Figure 7B, bottom).
These results clearly indicate that intracellular transportation of the truncated rXIIIB to the Golgi was impaired and it was stacked within the ER.
In the present study, 2 Japanese patients were newly diagnosed as having XIIIB deficiency. At this writing, as few as 5 cases of XIIIB deficiency have been identified in the world and these were found only in Italy and Japan. In contrast, more than 200 cases of XIIIA deficiency have been reported, and they were distributed worldwide. In general, XIIIA deficiency shows a nearly complete loss of the catalytic XIIIA protein in plasma, which is caused by defects in the XIIIA gene.14 On the other hand, the catalytic XIIIA in plasma significantly decreased but remained at about 5% to 40% of normal in some cases of XIIIB deficiency. Therefore, the clinical symptoms of XIIIB deficiency may be milder, and perhaps XIIIB deficiency is more difficult to discover than XIIIA deficiency. There may be many more undiagnosed cases of XIIIB deficiency in the world today; nevertheless, it appears to be an extremely rare condition. The ag A novel mutation, a deletion of guanosine in exon IX (delG), was
identified in one patient (patient 4) heterozygously as a second
mutation. As a result of this delG mutation, a premature stop codon
would be expected to be introduced in the XIIIB gene with a
frameshift, resulting in the production of a truncated XIIIB (Figure
8). The resulting truncated XIIIB will
then consist of 489 amino acids containing 7 normal tandem repeats
called "Sushi domains" and one partial Sushi domain to which 10 extra amino acids attached (Figure 8B, bold). A truncated rXIIIB of
about 60 kd was actually synthesized in the mutant rXIIIB-transfected cells, as shown by the current expression studies. Pulse-chase experiments and immunocytochemical staining of the cells bearing the
delG XIIIB mutant clearly demonstrated impaired intracellular transportation of the truncated rXIIIB from the ER to the Golgi, and
the consequent impaired secretion. Thus, it is noteworthy that
truncation as well as amino acid substitution of XIIIB results in the
same severe disorder.10
There has been no information on the exact tertiary structure of XIIIB to date, because XIIIB is not readily crystallized mainly because of the heterogeneity in its carbohydrate chains. Therefore, it is not possible to predict likely effects of this delG mutation on the structure of the XIIIB molecule by molecular modeling, as was done for XIIIA.15,16 As discussed above, the mutant XIIIB has 7 intact Sushi domains. However, there must be some conformational change, at least in its eighth Sushi domain because it is truncated in the middle of the domain and extends the length of 10 entirely new amino acids. Moreover, an additional cysteine residue is introduced in this region of the mutant molecule and remains unpaired. These structural changes would lead to misfolding of the abnormal XIIIB molecule at the C-terminal region, and possibly affect the entire structure of XIIIB. The truncated and misfolded mutant XIIIB might then be trapped by some ER proteins like chaperones, which would prevent its transportation from the ER to the Golgi.17 This mechanism is called ER retention.18 In a cultured mammalian cell line, the delG mutation of the XIIIB gene resulted in impaired intracellular transportation of the truncated rXIIIB from the ER to the Golgi. Accordingly, the transportation of the truncated XIIIB must also be impaired in vivo, more specifically in the liver of patient 4 because XIIIB is synthesized by hepatocytes.19,20 Immunoblot analysis of the patient's plasma indicated that XIIIA was also significantly reduced (Figure 1). No mutations of the XIIIA gene, however, were identified in patient 4 by sequencing analysis. Accordingly, it is highly likely that the lack of plasma XIIIB resulted in a secondary loss of XIIIA in the patient's plasma because of the innate instability of the XIIIA molecule. The results obtained by the current studies strengthen our previous conclusion that genetic defects of XIIIB are the basis for the former type I factor XIII deficiency.
The authors thank Mr. T. Tamura for sequencing these patients' XIIIA gene, Dr H. Kaetsu for providing purified plasma XIII and XIIIB, Drs T. Izumi and F. Tokunaga for helpful discussions, Dr T. Shiraishi for making a confocal microscope available, and Ms L. Boba for her help in preparation of the manuscript.
Submitted August 21, 2000; accepted January 9, 2001.
Supported in part by research grants from the Ministry of Education, Science and Culture, Japan (11470205,11770071), the Ministry of Health and Welfare, the Uehara Memorial Foundation (Japan), and the Ryoichi Naito Foundation for Medical Research (Japan).
The first and second authors contributed equally to the completion of this study.
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.
Reprints: Akitada Ichinose, Department of Molecular Patho-Biochemistry and Patho-Biology, Yamagata University School of Medicine, Iida-Nishi 2-2-2, Yamagata 990-9585 Japan; e-mail: aichinos{at}med.id.yamagata-u.ac.jp.
1. Duckert F. Documentation of the plasma factor XIII deficiency in man. Ann N Y Acad Sci. 1972;202:190-199[Medline] [Order article via Infotrieve]. 2. Lorand L, Losowsky MS, Miloszewski KJ. Human factor XIII: fibrin-stabilizing factor. Prog Hemost Thromb. 1980;5:245-290[Medline] [Order article via Infotrieve]. 3. Ichinose A. The physiology and biochemistry of factor XIII. In: Bloom AL,Forbes CD,Thomas DP,Tuddenham EGD, eds. Haemostasis and Thrombosis. Vol 1. 3rd ed. Edinburgh, UK: Churchill Livingstone; 1994:531-546. 4. Girolami A, Burul A, Fabris F, Cappellato G, Betterle C. Studies on factor XIII antigen in congenital factor XIII deficiency: a tentative classification of the disease in two groups. Folia Haematol. 1978;105:131-141. 5. Saito M, Asakura H, Yoshida T, et al. A familial factor XIII subunit B deficiency. Br J Haematol. 1990;74:290-294[Medline] [Order article via Infotrieve]. 6. Capellato MG, Lazzaro AR, Marafioti F, Polato G, Girolami A. A new family with congenital factor XIII deficiency showing a deficit of both subunit A and B: type I factor XIII deficiency. Haematologia. 1987;20:179-187[Medline] [Order article via Infotrieve]. 7. Rodeghiero F, Tosetto A, Di Bona E, Castaman G. Clinical pharmacokinetics of a placenta- derived factor XIII concentrate in type I and type II factor XIII deficiency. Am J Hematol. 1991;36:30-34[Medline] [Order article via Infotrieve]. 8. Ichinose A, Izumi T, Hashiguchi T. The normal and abnormal genes of the a and b subunits in coagulation factor XIII. Semin Thromb Hemost. 1996;22:385-391[Medline] [Order article via Infotrieve].
9.
Hashiguchi T, Saito M, Morishita E, Matsuda T, Ichinose A.
Two genetic defects in a patient with complete deficiency of the b-subunit for coagulation factor XIII.
Blood.
1993;82:145-150 10. Hashiguchi T, Ichinose A. Molecular and cellular basis of deficiency of the b subunit for factor XIII secondary to a Cys430-Phe mutation in the seventh Sushi domain. J Clin Invest. 1995;95:1002-1008.
11.
Izumi T, Hashiguchi T, Castaman G, et al.
Type I factor XIII deficiency is caused by a genetic defect of its b subunit: insertion of triplet AAC in exon III leads to premature termination in the second Sushi domain.
Blood.
1996;87:2769-2774 12. Souri M, Izumi T, Higashi Y, Girolami A, Ichinose A. A founder effect is proposed for factor XIII B subunit deficiency caused by the insertion of triplet AAC in exon III encoding the second Sushi domain. Thromb Haemost. 1998;80:211-213[Medline] [Order article via Infotrieve].
13.
Kaetsu H, Hashiguchi T, Foster D, Ichinose A.
Expression and release of the a and b subunits for human coagulation factor XIII in baby hamster kidney (BHK) cells.
J Biochem.
1996;119:961-969 14. Ichinose A, Souri M, Izumi T, Takahashi N. Molecular and genetic mechanisms of factor XIII A subunit deficiency. Semin Thromb Hemost. 2000;26:5-10[Medline] [Order article via Infotrieve].
15.
Takahashi N, Tsukamoto H, Umeyama H, Castaman G, Rodeghiero F, Ichinose A.
Molecular mechanisms of type II factor XIII deficiency: novel Gly562-Arg mutation and C-terminal truncation of the A subunit cause factor XIII deficiency as characterized in a mammalian expression system.
Blood.
1998;91:2830-2838 16. Ichinose A, Tsukamoto H, Izumi T, et al. Arg260-Cys mutation in severe factor XIII deficiency: conformational change of the A subunit is predicted by molecular modelling and mechanics. Br J Haematol. 1998;101:264-272[CrossRef][Medline] [Order article via Infotrieve]. 17. Klausner RD, Sitia R. Protein degradation in the endoplasmic reticulum. Cell. 1990;62:611-614[CrossRef][Medline] [Order article via Infotrieve].
18.
Halaban R, Svedine S, Cheng E, Smicun Y, Aron R, Hebert DN.
Endoplasmic reticulum retention is a common defect associated with tyrosinase-negative albinism.
Proc Natl Acad Sci U S A.
2000;97:5889-5894 19. Ichinose A, McMullen BA, Fujikawa K, Davie EW. Amino acid sequence of the b subunit of human factor XIII, a protein composed of ten repetitive segments. Biochemistry. 1986;25:4633-4638[CrossRef][Medline] [Order article via Infotrieve].
20.
Nagy JA, Henriksson P, McDonagh J.
Biosynthesis of factor XIII B subunit by human hepatoma cell lines.
Blood.
1986;68:1272-1279
© 2001 by The American Society of Hematology.
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
|
 |
|
  S. E. Iismaa, B. M. Mearns, L. Lorand, and R. M. Graham Transglutaminases and Disease: Lessons From Genetically Engineered Mouse Models and Inherited Disorders Physiol Rev, July 1, 2009; 89(3): 991 - 1023. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Ajzner, A. Schlammadinger, A. Kerenyi, Z. Bereczky, E. Katona, G. Haramura, Z. Boda, and L. Muszbek Severe bleeding complications caused by an autoantibody against the B subunit of plasma factor XIII: a novel form of acquired factor XIII deficiency Blood, January 15, 2009; 113(3): 723 - 725. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. E. Lovejoy, T. C. Reynolds, J. E. Visich, M. D. Butine, G. Young, M. A. Belvedere, R. C. Blain, S. M. Pederson, L. M. Ishak, and D. J. Nugent Safety and pharmacokinetics of recombinant factor XIII-A2 administration in patients with congenital factor XIII deficiency Blood, July 1, 2006; 108(1): 57 - 62. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Koseki-Kuno, M. Yamakawa, G. Dickneite, and A. Ichinose Factor XIII A subunit-deficient mice developed severe uterine bleeding events and subsequent spontaneous miscarriages Blood, December 15, 2003; 102(13): 4410 - 4412. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
 |
 |
|||||||
 |
 |
 |
 |
 |
 |
 |
 |
||
| Copyright © 2001 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||
Top
Abstract
Introduction
2-plasmin inhibitor, and fibrin and fibronectin. These
reactions increase the mechanical strength of the fibrin clot and its
resistance to proteolytic degradation, and enhance the assembly of the
extracellular matrix. Accordingly, deficiency of factor XIII results in
defective cross-linking reactions.
-mercaptoethanol (
