A Novel Polymorphism in Glycoprotein IV (Replacement of Proline-90 by Serine) Predominates in Subjects with Platelet GPIV Deficiency

Summary

To clarify the molecular basis of the deficiency of glycoprotein IV (GPIV) of the platelet surface, we analyzed GPIV cDNA synthesized from platelet RNA of five unrelated Japanese subjects whose platelets did not express GPIV.

We confirmed the presence of normal-sized GPIV mRNA in platelets from subjects with GPIV deficiency. The sequence of platelet GPIV cDNA from GPIV deficient subject showed three differences when compared with the published sequence; 1) a replacement of a ⁴⁷⁸CCT codon for proline-90 by TCT for serine, 2) a four-base insertion in the 3′-noncoding region, and 3) a substitution of A for ⁷⁹C in the 5′-noncoding region. The replacement of Pro⁹⁰ by Ser predominates in subjects with GPIV deficiency; that is, four out of five platelets with GPIV deficiency contained GPIV mRNA encoding GPIV^Ser-90, while all platelets from 17 GPIV positive subjects had GPIV mRNA encoding GPIV^Pro-90. The sequence of platelet GPIV cDNA which did not encode GPIV^Ser-90 from a subject with GPIV deficiency revealed no abnormality in the coding region. The four-base insertion in the 3′-noncoding region and the substitution of A for ⁷⁹C in the 5′-noncoding region seems to be unrelated to the expression of GPIV.

The substitution of Ser for Pro⁹⁰ might alter the GPIV structure or impair GPIV biosynthesis, resulting in a lack of detectable GPIV. This hypothesis remains to be tested.

• References

• 1 Tandon NN, Lipsky RH, Burgess WH, Jamieson GA. Isolation and characterization of platelet glycoprotein IV (CD36). J Biol Chem 1989; 264: 7570-7575
• 2 Knowles II DM, Tolidjian B, Marboe C, D'Agati V, Grimes M, Chess L. Monoclonal anti-human monocyte antibodies OKM1 and OKM5 possess distinctive tissue distribution including differential reactivity with vascular endothelium. J Immunol 1984; 132: 2170-2173
• 3 Barnwell JW, Ockenhouse CF, Knowles II DM. Monoclonal anti body OKM5 inhibits the in vitro binding of Plasmodium falciparuminfected erythrocytes to monocytes, endothelial, and C32 melanoma cells. J Immunol 1985; 135: 3494-3497
• 4 Greenwalt DE, Watt KWK, So OY, Jiwani N. PAS IV, an integral membrane protein of mammary epithelial cells, is related to platelet and endothelial cell CD36 (GPIV). Biochemistry 1990; 29: 7054-7059
• 5 Tandon NN, Kralisz U, Jamieson GA. Identification of GPIV (CD36) as a primary receptor for platelet-collagen adhesion. J Biol Chem 1989; 264: 7576-7583
• 6 Ockenhouse CF, Tandon NN, Magowan C, Jamieson GA, Chulay J. Identification of a platelet membrane glycoprotein as a falciparum malaria sequestration receptor. Science 1989; 243: 1469-1471
• 7 Oquendo P, Hundt E, Lawler J, Seed B. CD36 directly mediates cytoadherence of Plasmodium falciparum parasitized erythrocytes. Cell 1989; 58: 95-101
• 8 Ikeda H, Mitani T, Ohnuma M, Haga H, Ohtzuka S, Kato T, Nakase T, Sekiguchi S. A new platelet-specific antigen, Nak^a, involved in the refractoriness of HLA-matched platelet transfusion. Vox Sang 1989; 57: 213-217
• 9 Tomiyama Y, Take H, Ikeda H, Mitani T, Furubayashi T, Mizutani H, Yamamoto N, Tandon NN, Sekiguchi S, Jamieson GA, Kurata Y, Yonezawa T, Tarui S. Identification of the platelet-specific alloantigen, Nak^a, on platelet membrane glycoprotein IV. Blood 1990; 75: 684-687
• 10 Yamamoto N, Ikeda H, Tandon NN, Herman J, Tomiyama Y, Mitani T, Sekiguchi S, Lipsky R, Kralisz U, Jamieson GA. A platelet membrane glycoprotein (GP) deficiency in healthy blood donors: Nak^a- platelet lack detectable GPIV (CD36). Blood 1990; 76: 1698-1703
• 11 Kashiwagi H, Honda S, Take H, Mizutani H, Imai Y, Furubayashi T, Tomiyama Y, Kurata Y, Yonezawa T. Presence of the whole coding region of GPIV mRNA in Nak^a-negative platelets. Int J Hematol. (in press)
• 12 Lipsky RH, Sobieski DA, Tandon NN, Herman J, Ikeda H, Jamieson GA. Detection of GPIV (CD36) mRNA in Nak^a- platelets. Thromb Haemostas 1991; 65: 456-457
• 13 Kashiwagi H, Honda S, Take H, Mizutani H, Imai Y, Tomiyama Y, Kurata Y, Yonezawa T. Detection of Nak^a antigen and GPIV (CD36) mRNA in monocytes of Nak^a-negative subjects. Thromb Haemostas 1992; 67: 384-385
• 14 Takahashi TA, Hosoda S, Sekiguchi S. Cytospin method for the determination of residual leukocytes in leukocyte-depleted platelet concentrates. Jpn J Transfusion Med 1989; 35: 497-503
• 15 Knowles II DM, Tolidjian B, Marboe C, D'Agati V, Grimes M, Chess L. Monoclonal anti-human monocyte antibody OKM1 and OKM5 posses distinctive tissue distributions including differential reactivity with vascular endothelium. J Immunol 1984; 132: 2170-2173
• 16 Maniatis T, Fritsch EF, Sambrook J. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory; 1989: 196.
• 17 Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 1983; 132: 6-13
• 18 Sanger F, Nicken S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 1977; 74: 5463-5467
• 19 Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 1988; 239: 487-494
• 20 Lindeman R, Volpato SPHF, Trent RJ. Polymerase chain reacion (PCR) mutagenesis enabling rapid non-radioactive detection of common β-thalassaemia mutations in Mediterraneans. Br J Haematol 1991; 78: 100-104
• 21 Liebhaber SA, Kan YW. α-Thalassemia caused by an unstable α-globin mutant. J Clin Invest 1983; 71: 461-466
• 22 Mori Y, Takeda K, Charbonneau M, Refetoff S. Replacement of Leu²²⁷ by Pro in thyroxine-binding globulin (TBG) is associated with complete TBG deficiency in three of eight families with this inherited defect. J Clin Endocrinol Metab 1990; 70: 804-809
• 23 Alber T, Bell JA, Dao-Pin S, Nicholson H, Wozniak JA, Cook S, Matthews BW. Replacement of Pro⁸⁶ in phage T4 lysozyme extend an α-helix but do not alter protein stability. Science 1988; 239: 631-635
• 24 Brandts JF, Halvorson HR, Brennan M. Consideration of possibility that the slow step in protein denaturation reaction is due to cis-trans isomerism of proline residues. Biochemistry 1975; 14: 4953-4963
• 25 Stellwagen E. Proline peptide isomerization and the reactivation of denatured enzymes. J Mol Biol 1979; 135: 217-229