Heterogenous Expression of Glycoprotein lb, IX and V in Platelets from Two Patients with Bernard-Soulier Syndrome Caused by Different Genetic Abnormalities

Summary

Bernard-Soulier syndrome (BSS) is a rare inherited bleeding disorder, which is caused by deficiency or decrease of the platelet GPIb/IX/V complex. Analysis of two patients with BSS by How cytometry of the blood revealed different expression patterns of the components of the GPIb/IX/V complex. In case 1, GPIX was completely absent but residual amounts of GPIbα and GPV were detectable; in case 2, GPIbα was completely absent. We amplified the coding regions of GPIbα, GPIbß, GPV, and GPIX from the patients’ genomic DNA with the polymerase chain reaction (PCR) and sequenced the PCR products. In case 1, we identified a point mutation in the GPIX coding region that changes the codon for tryptophan-126 (TGG) to a nonsense codon (TGA). In case 2, we found a deletion of nucleotide within seven adenine repeats at the position of 1932 to 1938 in the coding region of GPIbα, which causes a frame shift that results in 58 altered amino acids and a premature stop codon. These genetic changes alter the transmembrane domain of GPIX or GPIbα and, therefore, would prevent proper insertion of the proteins in the plasma membrane. Thus, abnormality of a single component protein (GPIX or GPIbα) alters the assembly of the GPIb/IX/V complex and causes heterogenous surface expression of GPIbα, GPV and GPIX.

• References

• 1 Berndt MC, Fournier DJ, Castaldi PA. Bernard-Soulier syndrome. Baillieres Clin Haematol 1989; 2: 585-607
• 2 Nurden AT, Caen JP. Specific roles for platelet surface glycoproteins in platelet function. Nature 1975; 255: 720-722
• 3 Berndt MC, Gregory C, Chong BH, Zola H, Casyaldi PA. Additional glycoprotein delects in Bernard-Soulier’s syndrome: confirmation of genetic basis by parental analysis. Blood 1985; 62: 800-807
• 4 Roth GJ. Developing relationships: arterial platelet adhesion, glycoprotein lb, and leucine-rich glycoproteins. Blood 1991; 77: 5-19
• 5 Wieki AN, Clemetson KJ. Structure and function of platelet membrane glycoproteins lb and V. Effects of leukocyte elaslase and other proteases on platelets response to von Willebrand factor and thrombin Eur J Bioehcm 1985; 153: 1-11
• 6 Yamamoto K, Yamamoto N, Kitagawa H, Tanouc 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
• 7 Du X, Beutler L, Ruan C, Castakli PA, Berndt MC. Glycoprotein lb and glycoprotein IX are fully complexed in the intact platelet membrane. Blood 1987; 69: 1524-1527
• 8 Bennett JS. The molecular biology of platelet membrane proteins. Seminars in Haemat 1990; 27: 186-204
• 9 Modderman PW, Admirral LG, Sonnenberg A, von dem Borne AB G K r. Glycoprotein V and Ib-IX form a noncovalent complex in the platelet membrane. J Biol Chem 1992; 267: 364-369
• 10 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
• 11 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 lb. Blood 1990; 75: 2349-2356
• 12 Lopez JA, Chung DW, Fujikawa K, Hagen FS, Papayannopoulou T, Roth GJ. Cloning of the a chain of human platelet glycoprotein lb: a transmembrane protein with homology to leucine-rich α₂-glycoprotein. Proc Natl Acad Sci USA 1987; 84: 5615-5619
• 13 Wenger RH, Kieffer N, Wicki AN, Clemetson KJ. Structure of the human blood platelet membrane glycoprotein Ibα gene. Biochem Biophys Res Commun 1988; 156: 389-395
• 14 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 GPIbα gene to chromosome 22. J Clin Invest 1994; 93: 2417-2424
• 15 Yagi M, Edelhoff S, Disteche CM, Roth GJ. Structural characterization and chromosomal location of the gene encoding human platelet glycoprotein Ibβ. J Biol Chem 1994; 269: 17424-17427
• 16 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
• 17 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
• 18 Hickey MJ, Hagen FS, Yagi M, Roth GJ. Human platelet glycoprotein V: Characterization of the polypeptide and the related Ib-V-IX receptor system of adhesive, leucine-rich glycoproteins. Proc Natl Acad Sci USA 1993; 90: 8327-8331
• 19 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
• 20 Ware J, Russell SR, Vincente V, Scharf RE, Tomer A, McMillan R, Ruggeri ZM. Nonsense mutation in the glycoprotein Iba coding sequence associaled with Bernard Soulier syndrome. Proc Nall Acad Sci USA 1960; 87: 2026-2030
• 21 Miller JL, Lyle VA, Cunningham D. Mutation of leucine 57 to phenylalanine in a platelet glycoprotein lbα leucine tandem repeal occurring in patients with an autosomal dominant variant of Bernard Soulier disease. Blood 1992; 79: 439-446
• 22 Ware J, Russell SR, Marchesc P, Murata M, Mazzucalo M, Marco LD, Ruggeri ZM. Point mulalion in a leucine-rich repeal of platelet glycoprotein Iba resulting in the Bernard Soulier syndrome. J Clin Invest 1993; 92: 1213-1220
• 23 Kunishima S, Miura H, Fukutani H, Yoshida H, Osumi K, Kobayashi S, Ohno R, Naoe T. Bernard Soulier syndrome Kagoshima: Ser 444 > stop mutation of glycoprotein (GP) Ibα resulting in circulating truncated GPIbα and surface expression of GPIbβi and GPIX. Blood 1994; 84: 3356-3362
• 24 Simsek S, Noris P, Lozano M, Pico M, von dem Borne AB G K r, Ribera A, Gallardo D. Cys209 Ser mutation in the platelet membrane glycoprotein Ibα gene is associated with Bernard-Soulier syndrome. Brit J Haematol 1994; 88: 839-844
• 25 Simsek S, Admiraal LG, Modderman PW, van der School CE, von dem Borne AB G K r. Identification of a homozygous single base pair deletion in the gene coding for the human platelet glycoprotein Ibα causing Bernard-Soulier syndrome. Thromb Haemost 1994; 72: 444-449
• 26 de la Salle C, Baas MJ, Lanza F, Schwartz A, Hanau D, Chevalier J, Gachet C, Briquel MB, Cazenave JP. A three-base deletion removing a leucine residue in a leucine-rich repeat of platelet glycoprotein Ibα associated with a variant of Bernard-Soulier syndrome (Nancy I). Brit J Haematol 1995; 89: 386-396
• 27 Wright SD, Michaelides K, Johnson DJ D, West NC, Tuddenham EG D. Double heterozygosity for mutations in the platelet glycoprotein IX gene in the three siblings with Bernard-Soulier syndrome. Blood 1993; 81: 2339-2347
• 28 Clemetson JM, Kyrle PA, Brenner B, Clemetson KJ. Variant Bernard-Soulier syndrome associated with a homozygous mutation in the leucine-rich domain of glycoprotein IX. Blood 1994; 84: 1124-1131
• 29 Arai M, Yamamoto N, Akamatsu N, Suzuki H, Yamaguchi A, Nishida Y, Fukutake K, Tanoue K. Substantial expression of glycoproteins IX and V on the platelet surface from a patient with Bernard-Soulier syndrome. Br J Haematol 1994; 87: 185-188
• 30 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
• 31 Blin N, Stafford DW. Isolation of high molecular weight DNA. Nucleic Acids Res 1976; 3: 2303
• 32 Lopez JA, Ludwig EH, McCarthy BJ. Polymorphism of human glycoprotein Ibα 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
• 33 Lopez JA, Leung B, Reynolds CC, Li CQ, Fox JE B. 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
• 34 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
• 35 George JN, Caen JP, Nurden AT. Glanzmann’s thrombasthenia: the spectrum of clinical disease. Blood 1990; 75: 1383-1395
• 36 Arnaout MA. Structure and function of the leucocyte adhesion molecules CD11/CD18. Blood 1990; 75: 1037-1050