A Point Mutation in Glycoprotein IX Coding Sequence (Cys73(TGT) to Tyr(TAT)) Causes Impaired Surface Expression of GPIb/IX/V Complex in Two Families with Bernard-Souiier Syndrome

Masaaki Noda; Kingo Fujimura; Toshiro Takafuta; Takeshi Shimomura; Teruhisa Fujii; Shinya Katsutani; Tetsuro Fujimoto; Atsushi Kuramoto; Tomio Yamazaki; Toshihiro Mochizuki; Miwako Matsuzaki; Masayuki Sano

doi:10.1055/s-0038-1650678

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1996; 76(06): 0874-0878
DOI: 10.1055/s-0038-1650678

Original Article

Schattauer GmbH Stuttgart

A Point Mutation in Glycoprotein IX Coding Sequence (Cys⁷³(TGT) to Tyr(TAT)) Causes Impaired Surface Expression of GPIb/IX/V Complex in Two Families with Bernard-Souiier Syndrome

Masaaki Noda

¹The Department of Hematology and Oncology, Division of Clinical Research, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima

,

Kingo Fujimura

¹The Department of Hematology and Oncology, Division of Clinical Research, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima

,

Toshiro Takafuta

¹The Department of Hematology and Oncology, Division of Clinical Research, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima

,

Takeshi Shimomura

¹The Department of Hematology and Oncology, Division of Clinical Research, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima

,

Teruhisa Fujii

¹The Department of Hematology and Oncology, Division of Clinical Research, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima

,

Shinya Katsutani

¹The Department of Hematology and Oncology, Division of Clinical Research, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima

,

Tetsuro Fujimoto

¹The Department of Hematology and Oncology, Division of Clinical Research, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima

,

Atsushi Kuramoto

¹The Department of Hematology and Oncology, Division of Clinical Research, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima

,

Tomio Yamazaki

²First Department of Internal Medicine, Nagoya University School of Medicine, Aichi

,

Toshihiro Mochizuki

³Department of Internal Medicine, Shizuoka City Hospital, Shizuoka

,

Miwako Matsuzaki

⁴Department of Internal Medicine, Saga College of Medicine, Saga, Japan

,

Masayuki Sano

⁴Department of Internal Medicine, Saga College of Medicine, Saga, Japan

› Institutsangaben

Abstract

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Summary

Bernard-Soulier syndrome (BSS) is a rare inherited bleeding disorder which is caused by abnormal expression or function of the glycoprotein (GP) Ib/IX/V complex, a platelet major receptor for von Wille-brand factor. We studied four BSS patients in two unrelated families in which the same and novel mutation was found. Flow cytometric analysis showed that GPIX was completely absent but residual amounts of GPIbα and GPV were detectable in these patients. We analyzed all coding regions of GPIbα, GPIbβ, GPV and GPIX which were amplified from the patients’ genomic DNA by the polymerase chain reaction (PCR). In all four cases, we identified a point mutation in the GPIX coding region that changes the codon for cysteine 73 (TGT) to a codon for tyrosine (TAT). Furthermore, we confirmed by a transient expression study that the mutation caused the loss of adequate surface expression of GPIX. Since cysteine might be important for the secondary structure, this mutation of GPIX gene would lead to a dramatic conformational change of GPIX protein, resulting in the reduced surface expression. We concluded that this novel point mutation of the GPIX gene was responsible for BSS in these families

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Referenzen

References
1 Roth GJ. Developing relationships: arterial platelet adhesion, glycoprotein Ib, and leucine-rich glycoproteins. Blood 1991; 77: 5-19
2 Ruggeri ZM, De Marco L, Gatti L, Bader R, Montgomery RR. Platelets have more than one binding site for von Willebrand factor. J Clin Invest 1983; 72: 1-12
3 Wicki AN, Clemetson KJ. Structure and function of platelet membrane glycoproteins lb and V. Effects of leukocyte elastase and other proteases on platelets response to von Willebrand factor and thrombin. Eur J Biochem 1985; 153: 1-11
4 Yamamoto K, Yamamoto N, Kitagawa H, Tanoue K, Kosaki G, Yamazaki H. Localization of a thrombin-binding site on human platelet membrane glycoprotein lb determined by a monoclonal antibody. Thromb Haemost 1986; 55: 162-167
5 Du X, Beutler L, Ruan C, Castaldi PA, Bemdt MC. Glycoprotein lb and glycoprotein IX are fully complexed in the intact platelet membrane. Blood 1987; 69: 1524-1527
6 Modderman PW, Admirral LG, Sonnenberg A, von dem Borne AEGKr. Glycoprotein V and Ib-IX form a noncovalent complex in the platelet membrane. J Biol Chem 1992; 267: 364-369
7 Lopez JA, Chung DW, Fujikawa K, Hagen FS, Davie EW, Roth GJ. The α and β chains of human platelet glycoprotein lb are both transmembrane proteins containing a leucine-rich amino acid sequence. Proc Natl Acad Sci USA 1988; 85: 2135-2139
8 Shimomura T, Fujimura K, Maehama S, Takemoto M, Oda K, Fujimoto T, Oyama R, Suzuki M, Ichihara-Tanaka K, Titani K, Kuramoto A. Rapid purification and characterization of human platelet glycoprotein V: the amino acid sequence contains leucine-rich repetitive modules as in glycoprotein Ib. Blood 1990; 75: 2349-2356
9 Bemdt MC, Fournier DJ, Castaldi PA. Bemard-Soulier syndrome. Baillierres Clin Haematol 1989; 2: 585-607
10 Nurden AT, Caen JP. Specific roles for platelet surface glycoproteins in platelet function. Nature 1975; 255: 720-722
11 Bemdt MC, Gregory C, Chong BH, Zola H, Castaldi PA. Asditional glycoprotein defects in Bemard-Soulier’s syndrome: confirmation of genetic basis by parental analysis. Blood 1983; 62: 800-807
12 Wenger RH, Kieffer N, Wicki AN, Clemetson KJ. Structure of the human blood platelet membrane glycoprotein Iba gene. Biochem Biophys Res Commun 1988; 156: 389-395
13 Kelly MD, Essex DW, Shapiro SS, Meloni FJ, Druck T, Huebner K, Konkle BA. Complementary DNA cloning of the alternatively expressed endothelial cell glycoprotein Ibβ and localization of the GPIbβ gene to chromosome 22. J Clin Invest 1994; 93: 2417-2424
14 Hickey MJ, Williams SA, Roth GJ. Human platelet glycoprotein IX: an adhesive prototype of leucine-rich glycoproteins with flank-center-flank structures. Proc Natl Acad Sci USA 1989; 86: 6773-6777
15 Hickey MJ, Deaven LL, Roth GJ. Human platelet glycoprotein IX: Characterization of cDNA and localization of the gene to chromosome 3. Fed Eur Biochem Soc 1990; 274: 189-192
16 Lanza F, Morales M, de La Salle C, Cazenave JP, Clemetson KJ, Shimomura T, Phillips DR. Cloning and characterization of the gene encoding the human platelet glycoprotein V. J Biol Chem 1993; 268: 20801-20807
17 Ware J, Russell SR, Vincente V, Scharf RE, Tomer A, McMillan R, Ruggeri ZM. Nonsense mutation in the glycoprotein Iba coding sequence associated with Bemard-Soulier syndrome. Proc Natl Acad Sci USA 1990; 87: 2026-2030
18 Miller JL, Lyle VA, Cunningham D. Mutation of leucine-57 to phenylalanine in a platelet glycoprotein Iba leucine tandem repeat occurring in patients with an autosomal dominant variant of Bernard-Soulier disease. Blood 1992; 79: 439-446
19 Ware J, Russell SR, Marchese P, Murata M, Mazzucato M, Marco LD, Ruggeri ZM. Point mutation in a leucine-rich repeat of platelet glycoprotein Iba resulting in the Bemard-Soulier syndrome. J Clin Invest 1993; 92: 1213-1220
20 Kunishima S, Miura H, Fukutani H, Yoshida H, Osumi K, Kobayashi S, Ohno R, Naoe T. Bemard-Soulier syndrome Kagoshima: Ser 444 →stop mutation of glycoprotein (GP) Ibα resulting in circulating truncated GPIba and surface expression of GPIbβ and GPIX. Blood 1994; 84: 3356-3362
21 Simsek S, Noris P, Lozano M, Pico M, von dem Bome AEGKr, Ribera A, Gallardo D. Cys209 Ser mutation in the platelet membrane glycoprotein Iba gene is associated with Bemard-Soulier syndrome. Brit J Haematol 1994; 88: 839-844
22 Li C, Martin SE, Roth GJ. The genetic defect in two well-studied cases of Bemard-Soulier syndrome: A point mutation in the fifth leucine-rich repeat of platelet glycoprotein Ibα. Blood 1995; 86: 3805-3814
23 Simsek S, Admiraal LG, Modderman PW, van der Schoot CE, von dem Borne AEGKr. Identification of a homozygous single base pair deletion in the gene coding for the human platelet glycoprotein Iba causing Bernard-Soulier syndrome. Thromb Haemost 1994; 72: 444-449
24 de la Salle C, Baas MJ, Lanza F, Schwartz A, Hanau D, Chevalier J, Gachet C, Briquel ME, Cazenave JP. A three-base deletion removing a leucine residue in a leucine-rich repeat of platelet glycoprotein Iba associated with a varient of Bemard-Soulier syndrome (Nancy I). Brit J Haematol 1995; 89: 386-396
25 Noda M, Fujimura K, Takafuta T, Shimomura T, Fujimoto T, Yamamoto N, Tanoue K, Arai M, Suehiro A, Kakishita E, Shimasaki A, Kuramoto A. Heterogenous expression of glycoprotein Ib, IX and V in platelets from two patients with Bemard-Soulier syndrome caused by different genetic abnormalities. Thromb Haemost 1995; 74: 1411-1415
26 Wright SD, Michaelides K, Johnson DJD, West NC, Tuddenham EGD. Double heterozygosity for mutations in the platelet glycoprotein IX gene in the three siblings with Bemard-Soulier syndrome. Blood 1993; 81: 2339-2347
27 Clemetson JM, Kyrle PA, Brenner B, Clemetson KJ. Variant Bemard-Soulier syndrome associated with a homozygous mutation in the leucine-rich domain of glycoprotein IX. Blood 1994; 84: 1124-1131
28 Budarf ML, Konkle BA, Ludlow LB, Michaud D, Li M, Yamashiro DJ, MuGinn DM, Zackai EH, Driscoll DA. Identification of a patient with Bemard-Soulier syndrome and a deletion in the DiGeorge/Velo-cardio-facial chromosomal region in 22ql 1.2. Human Molecular Genetics 1995; 4: 763-766
29 Takafuta T, Fujimura K, Kawano H, Noda M, Fujimoto T, Oda K, Shimomura T, Kuramoto A. Expression of platelet membrane glycoprotein V in human megakaryocytes and megakaryocytic cell lines: a study using a novel monoclonal antibody against GPV. Thromb Haemost 1994; 72: 762-769
30 Sambrook J, Fritsch EF, Maniatis T. Isolation of high molecular weight DNA from mammalian cells. Molecular Cloning, 2nd ed 1989; 9.: 14-23
31 Higuchi R, Krummel B, Saiki RK. A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. Nucleic Acids Res 1988; 16: 7351-7367
32 Mizushima S, Nagata S. pBK-BOS, a powerful mammalian expression vector. Nucleic Acids Res 1990; 18: 5322
33 Fujimoto T, McEver RP. The cytoplasmic domain of P-selectin is phos-phorylated on serine and threonine residues. Blood 1993; 82: 1758-1766
34 Lopez JA, Ludwig EH, McCarthy BJ. Polymorphism of human glycoprotein Iba results from a variable number of tandem repeats of a 13-amino acid sequence in the mucin-like macroglycopeptide region. J Biol Chem 1992; 267: 10055-10061
35 Lopez JA. The platelet glycoprotein Ib-IX complex. Blood Coagul Fibrinol 1994; 5: 97-119
36 Sae-Tung G, Dong J, Lopez JA. Biosynthetic defect in platelet glycoprotein IX mutants associated with Bemard-Soulier syndrome. Blood 1996; 87: 1361-1367
37 Lopez JA, Leung B, Reynolds CC, Li CQ, Fox JEB. Efficient plasma membrane expression of a functional platelet glycoprotein Ib-IX complex requires the presence of its three subunits. J Biol Chem 1992; 267: 12851-12859
38 Meyer S, Kresbach G, Haring P, Schumpp-Vonach B, Clemetson KJ, Hadvary P, Steiner B. Expression and characterization of functionally active fragments of the platelet glycoprotein Ib-IX complex in mammalian cells. J Biol Chem 1993; 268: 20555-20562
39 Lopez JA, Weisman S, Sanan DA, Sih T, Chambers M, Li CQ. Glycoprotein Ibβ is the critical subunit linking GPIbα and GPIX in the GPIb-IX complex. J Biol Chem 1994; 269: 23716-23721
40 Meyer SC, Fox JE. Interaction of platelet glycoprotein V with glycoprotein Ib-IX regulates expression of the glycoproteins and binding of von Wille-brand factor to glycoprotein Ib-IX in transfected cells. J Biol Chem 1995; 270: 14693-14699