title: Human endothelial cells in culture and in vivo express on their surface all four components of the glycoprotein Ib/IX/V complex.
abstract: The platelet glycoprotein Ib (GpIb) complex is composed of four polypeptides: the disulfide-linked GpIb alpha and GpIb beta and the noncovalently associated GpIX and GpV. GpIb alpha contains binding sites for von Willebrand factor and for thrombin and mediates platelet adhesion to the subendothelium under conditions of high shear stress. We have previously shown the presence of GpIb alpha and GpIb beta mRNA and protein in cultured human umbilical vein endothelial cells (HUVECs) as well as the presence of GpIb alpha mRNA and protein in tonsillar endothelium. We, therefore, probed ECs for the presence of the other components of the GpIb/IX/V complex. We have identified the presence of GpIX and GpV mRNA in cultured HUVEC monolayers. The sequence of HUVEC GpIX cDNA was identical to the previously published human erythroleukemia (HEL) cell GpIX cDNA sequence. Two species of GpV mRNA, one of 3 kb and one of 4.4 kb, were found in HUVECs, whereas HEL cells displayed only the 4.4-kb species and the megakaryocytic cell line CHRF-288 contained only the 3-kb species. We previously showed that EC GpIb alpha protein is identical in molecular weight to platelet GpIb alpha. HUVEC GpIb beta, in contrast to its platelet counterpart, has a molecular weight of 50 kD and forms a correspondingly larger disulfide-bonded complex with EC GpIb alpha. The molecular weights of GpIX and GpV were 22 and 88 kD, respectively, identical to the corresponding platelet polypeptides. Furthermore, we have identified all four components of the complex in tonsillar vessels. Using flow cytometry, we have established that all four polypeptides of the GpIb/IX/V complex are expressed on the surface membranes of cultured HUVECs and adult aortic ECs. Furthermore, using two-color fluorescence, we have shown that all ECs expressing GpIb alpha also express GpIX and GpV on their surface. The ratio of GpIb alpha:GpIX:GpV is 1:1:0.5, which is identical to the ratio present in platelets. None of the polypeptides of the GpIb complex could be identified on the surface of human smooth muscle cells or lymphocytes. The presence of all members of the GpIb complex in the EC membrane suggests that this complex may play a role in endothelial function in vivo.
authors.affiliation: Cardeza Foundation for Hematologic Research and the Department of Medicine, Jefferson Medical College of Thomas Jefferson University, Philadelphia, PA 19107-5099, USA.
authors.forename: G
authors.initials: G
authors.lastname: Wu
authors.forename: D W
authors.initials: DW
authors.lastname: Essex
authors.forename: F J
authors.initials: FJ
authors.lastname: Meloni
authors.forename: T
authors.initials: T
authors.lastname: Takafuta
authors.forename: K
authors.initials: K
authors.lastname: Fujimura
authors.forename: B A
authors.initials: BA
authors.lastname: Konkle
authors.forename: S S
authors.initials: SS
authors.lastname: Shapiro
mesh_terms: D000328:Adult; D001011:Aorta / Q000166:cytology; D002478:Cells, Cultured; D004727:Endothelium / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism; D006801:Humans; D007231:Infant, Newborn; D008214:Lymphocytes / Q000737:chemistry; D008970:Molecular Weight; D009130:Muscle, Smooth / Q000737:chemistry; D009928:Organ Specificity; D014066:Palatine Tonsil / Q000166:cytology; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014471:Umbilical Veins / Q000166:cytology
chemical_list: D019038:Platelet Glycoprotein GPIb-IX Complex; D012333:RNA, Messenger