Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter June 1, 2005

Polymorphisms in the P-selectin (CD62P) and P-selectin glycoprotein ligand-1 (PSGL-1) genes and coronary heart disease

  • Peter Bugert , Marion Vosberg , Mathias Entelmann , Jürgen Jahn , Hugo A. Katus and Harald Klüter


P-selectin and its ligand, PSGL-1, are cell adhesion molecules that facilitate interaction of platelets, leukocytes and endothelial cells. Polymorphisms of these genes have been reported to be associated with coronary heart disease (CHD). In the present study, we characterized the entire coding regions of P-selectin and PSGL-1 genes in CHD patients and healthy controls. The 17 exons of the P-selectin gene and exon 2 of the PSGL-1 gene were screened for single nucleotide polymorphisms (SNPs) by exon re-sequencing in 88 CHD patients and 96 controls. For rapid genotyping of the SNPs we developed PCR techniques with sequence-specific primers (PCR-SSP). By using PCR-SSPs we genotyped 261 CHD patients and 214 controls for 5 SNPs in P-selectin and 2 SNPs in PSGL-1. In addition to the already described SNPs in P-selectin (S290N, N562D, V599L and T715P), we identified a novel SNP in exon 5 (V168M). The P-selectin 715P allele was more frequent among CHD patients with hypercholesterolemia compared to patients with normal cholesterol levels. A SNP (M62I) in the PSGL-1 gene was found close to the P-selectin binding site and the 62I allele revealed a higher prevalence in the control group indicating a protective effect of the mutation. The molecular characterization of P-selectin and PSGL-1 in a case-control study including CHD patients and healthy controls revealed evidence for association of the genes with development of the disease. However, the functional role of the gene variants should be elucidated by further experimental data.

Corresponding author: Dr. Peter Bugert, Institut für Transfusionsmedizin und Immunologie, DRK-Blutspendedienst Baden-Württemberg – Hessen gGmbH, Friedrich-Ebert-Straße 107, 68167 Mannheim, Germany. Phone: +49-621-3706-8122, Fax: +49-621-3706-876, E-mail:


1 Vestweber D, Blanks JE. Mechanisms that regulate the function of the selectins and their ligands. Physiol Rev 1999; 79:181–213.10.1152/physrev.1999.79.1.181Search in Google Scholar

2 Mayadas TN, Johnson RC, Rayburn H, Hynes RO, Wagner DD. Leukocyte rolling and extravasation are severely comprised in P-selectin-deficient mice. Cell 1993; 74:541–54.10.1016/0092-8674(93)80055-JSearch in Google Scholar

3 Dong ZM, Chapman SM, Brown AA, Frenette PS, Hynes RO, Wagner DD. The combined role of P and E selectins in atherosclerosis. J Clin Invest 1998; 102:145–52.10.1172/JCI3001Search in Google Scholar

4 Celi A, Lorenzet R, Furie B, Furi BC. Platelet-leukocyte-endothelial cell interaction on the blood vessel wall. Semin Hematol 1997; 34:327–35.Search in Google Scholar

5 Borges E, Eytner R, Moll T, Steegmaier M, Campbell MA, Ley K, et al. The P-selectin glycoprotein ligand-1 is important for recruitment of neutrophils into inflamed mouse peritoneum. Blood 1997; 90:1934–42.10.1182/blood.V90.5.1934Search in Google Scholar

6 Ramachandran V, Yago T, Epperson TK, Kobzdej MMA, Nollert MU, Cummings RD, et al. Dimerization of a selectin and its ligand stabilizes cell rolling and enhances tether strength in shear flow. Proc Natl Acad Sci USA 2001; 98:10166–71.10.1073/pnas.171248098Search in Google Scholar

7 Frenette PS, Denis CV, Weiss L, Jurk K, Subbarao S, Kehrel B, et al. P-selectin glycoprotein ligand 1 (PSGL-1) is expressed on platelets and can mediate platelet-endothelial interactions in vivo. J Exp Med 2000; 191:1413–22.10.1084/jem.191.8.1413Search in Google Scholar

8 Foxall C, Watson SR, Dowbenko D, Fennie C, Lasky LA, Kiso M, et al. The three members of selectin receptor family recognize a common carbohydrate epitope, the sialyl Lewis X oligosaccharide. J Cell Biol 1992; 117:895–902.10.1083/jcb.117.4.895Search in Google Scholar

9 Pouyani T, Seed B. PSGL-1 recognition of P-selectin is controlled by a tyrosine sulfation consensus at the PSGL-1 amino terminus. Cell 1995; 83:333–43.10.1016/0092-8674(95)90174-4Search in Google Scholar

10 Sako D, Comess KM, Barone KM, Camphausen RT, Cumming DA, Shaw GD. A sulfated peptide segment at the amino terminus of PSGL-1 is critical for P-selectin binding. Cell 1995; 83:323–31.10.1016/0092-8674(95)90173-6Search in Google Scholar

11 Wilkins PP, McEver RP, Cummings RD. Structures of the O-glycans on P-selectin glycoprotein ligand-1 from HL-60 cells. J Biol Chem 1996; 271:18732–42.10.1074/jbc.271.31.18732Search in Google Scholar PubMed

12 Herrmann S-M, Ricard S, Nicaud V, Mallet C, Evans A, Ruidavets J-B, et al. The P-selectin gene is highly polymorphic: reduced frequency of the Pro715 allele carriers in patients with myocardial infarction. Hum Mol Genet 1998; 7:1277–84.10.1093/hmg/7.8.1277Search in Google Scholar PubMed

13 Kee F, Morrison C, Evans EA, McCrum E, McMaster D, Dallongeville J, et al. Polymorphisms of the P-selectin gene and risk of myocardial infarction in men and women in the ECTIM study. Heart 2000; 84:548–52.10.1136/heart.84.5.548Search in Google Scholar PubMed PubMed Central

14 Afshar-Kharghan V, Diz-Küçükkaya R, Ludwig EH, Marian AJ, Lopez JA. Human polymorphism of P-selectin glycoprotein ligand 1 attributeable to variable numbers of tandem decameric repeats in the mucinlike region. Blood 2001; 97:3306–7.10.1182/blood.V97.10.3306Search in Google Scholar

15 Lozano ML, González-Conejero R, Corral J, Rivera J, Iniesta JA, Martinez C, et al. Polymorphisms of P-selectin glycoprotein ligand-1 are associated with neutrophil-platelet adhesion and with ischaemic cerebrovascular disease. Br J Haematol 2001; 115:969–76.10.1046/j.1365-2141.2001.03151.xSearch in Google Scholar PubMed

16 Tregouet DA, Barbaux S, Poirier O, Blankenberg S, Bickel C, Escolano S, et al. SELPLG gene polymorphisms in relation to plasma SELPLG levels and coronary artery disease. Ann Hum Genet 2003; 67:504–11.10.1046/j.1529-8817.2003.00053.xSearch in Google Scholar PubMed

17 Bugert P, Decker S, Klüter H. Exon-Amplification-Restriction-Ligation (EARL): an efficient strategy for direct sequencing of exons. Biotechniques 2001; 30:490–6.10.2144/01303bm05Search in Google Scholar PubMed

18 Bugert P, Lese A, Meckies J, Zieger W, Eichler H, Klüter H. Optimized sensitivity of allele-specific PCR for prenatal typing of human platelet alloantigen single nucleotide polymorphisms. Biotechniques 2003; 35:170–4.10.2144/03351md05Search in Google Scholar PubMed

19 Bugert P, Hoffmann MM, Winkelmann BR, Vosberg M, Jahn J, Entelmann M, et al. The variable number of tandem repeat polymorphism in the P-selectin glycoprotein ligand-1 (PSGL-1) gene is not associated with coronary heart disease. J Mol Med 2003; 81:495–501.10.1007/s00109-003-0459-2Search in Google Scholar PubMed

20 Revelle BM, Scott D, Kogan TP, Zheng J, Beck PJ. Structure-function analysis of P-selectin-sialyl Lewis X binding interactions. Mutagenic alteration of ligand binding specificity. J Biol Chem 1996; 271:4289–97.10.1074/jbc.271.8.4289Search in Google Scholar PubMed

21 Hirose M, Kawashima H, Miyasaka M. A functional epitope on P-selectin that supports binding of P-selectin to P-selectin glycoprotein ligand-1 but not to sialyl Lewis X oligosaccharides. Int Immunol 1998; 10:639–49.10.1093/intimm/10.5.639Search in Google Scholar PubMed

22 Revelle BM, Scott D, Beck PJ. Single amino acid residues in the E- and P-selectin epidermal growth factor domains can determine carbohydrate binding specificity. J Biol Chem 1996; 271:16160–70.10.1074/jbc.271.27.16160Search in Google Scholar PubMed

23 Tregouet DA, Barbaux S, Escolano S, Tahri N, Golmard J-L, Tiret L, et al. Specific haplotypes of the P-selectin gene are associated with myocardial infarction. Hum Mol Genet 2002; 11:2015–23.10.1093/hmg/11.17.2015Search in Google Scholar PubMed

24 Barbaux SC, Blankenberg S, Rupprecht HJ, Francomme C, Bickel C, Hafner G, et al. Association between P-selectin gene polymorphisms and soluble P-selectin levels and their relation to coronary artery disease. Arterioscler Thromb Vasc Biol 2001; 21:1668–73.10.1161/hq1001.097022Search in Google Scholar PubMed

25 Ikeda H, Nakayama H, Oda T, Kuwano K, Muraishi A, Sugi K, et al. Soluble form of P-selectin in patients with acute myocardial infarction. Coron Artery Dis 1994; 5:515–8.Search in Google Scholar

26 Ikeda H, Takajo Y, Ichiki K, Ueno T, Maki S, Noda T, et al. Increased soluble form of P-selectin in patients with unstable angina. Circulation 1995; 92:1693–6.10.1161/01.CIR.92.7.1693Search in Google Scholar PubMed

27 Davi G, Romano M, Mezzetti A, Procopio A, Iacobelli S, Antidormi T, et al. Increased levels of soluble P-selectin in hypercholesterolemic patients. Circulation 1998; 97:953–7.10.1161/01.CIR.97.10.953Search in Google Scholar

28 Tailor A, Granger DN. Hypercholesterolemia promotes P-selectin-dependent platelet-endothelial cell adhesion in postcapillary venules. Arterioscler Thromb Vasc Biol 2003; 23:675–80.10.1161/01.ATV.0000056742.97580.79Search in Google Scholar PubMed

29 Ishikawa M, Stokes KY, Zhang JH, Nanda A, Granger DN. Cerebral microvascular responses to hypercholesterolemia: roles of NADPH oxidase and P-selectin. Circ Res 2004; 94:239–44.10.1161/01.RES.0000111524.05779.60Search in Google Scholar PubMed

30 Merten M, Dong JF, Lopez JA, Thiagarajan P. Cholesterol sulfate: a new adhesive molecule for platelets. Circulation 2001; 103:2032–4.10.1161/01.CIR.103.16.2032Search in Google Scholar

31 Drayer NM, Lieberman S. Isolation of cholesterol sulfate from human aortas and adrenal tumors. J Clin Endocrinol Metab 1967; 27:136–9.10.1210/jcem-27-1-136Search in Google Scholar PubMed

32 Tamasawa N, Tamasawa A, Takebe K. Higher levels of plasma cholesterol sulfate in patients with liver cirrhosis and hypercholesterolemia. Lipids 1993; 28:833–6.10.1007/BF02536238Search in Google Scholar PubMed

33 Somers WS, Tang J, Shaw GD, Camphausen RT. Insights into the molecular basis of leukocyte tethering and rolling revealed by structures of P- and E-selectin bound to SLex and PSGL-1. Cell 2000; 103:467–79.10.1016/S0092-8674(00)00138-0Search in Google Scholar

Received: 2003-9-4
Accepted: 2004-6-29
Published Online: 2005-6-1
Published in Print: 2004-9-1

© Walter de Gruyter

Downloaded on 21.2.2024 from
Scroll to top button