title: The relative priority of prekallikrein and factors XI/XIa assembly on cultured endothelial cells.
abstract: Investigations determined the relative preference of prekallikrein (PK) or factor XI/XIa (FXI/FXIa) binding to endothelial cells (HUVECs). In microtiter plates, biotinylated high molecular weight kininogen (biotin-HK) or biotin-FXI binding to HUVEC monolayers or their matrix proteins, but not fibronectin-coated plastic microtiter plate wells, was specifically blocked by antibodies to each of the receptors of HK, uPAR, gC1qR, or cytokeratin 1. Fluorescein isothiocyanate (FITC)-PK specifically bound to HUVEC suspensions without added Zn2+, whereas FITC-FXI or -FXIa binding to HUVEC suspensions required 10 microM added Zn2+ to support specific binding. Plasma concentrations of FXI did not block FITC-PK binding to HUVECs in the absence or presence of 10 microM Zn2+. In the absence of HK, the level of FITC-FXI or -FXIa binding was half that seen in its presence. At physiologic concentrations, PK (450 nM) abolished FITC-FXI or -FXIa binding to HUVEC suspensions in the absence or presence of HK in the presence of 10 microM Zn2+. Released Zn2+ from 2-8 x 10(8) collagen-activated platelets/ml supported biotin-FXI binding to HUVEC monolayers, but platelet activation was not necessary to support biotin-PK binding to HUVECs. At physiologic concentrations, PK also abolished FXI binding to HUVECs in the presence of activated platelets, but FXI did not influence PK binding. PK in the presence or absence of HK preferentially bound to HUVECs over FXI or FXIa. Elevated Zn2+ concentrations are required for FXI but not PK binding, but the presence of physiologic concentrations of PK and HK also prevented FXI binding. PK preferential binding to endothelial cells contributes to their anticoagulant nature.
authors.affiliation: Department of Internal Medicine, Hematology and Oncology Division, the University of Michigan, Ann Arbor, Michigan 48109-0640, USA.
authors.forename: Fakhri
authors.initials: F
authors.lastname: Mahdi
authors.forename: Zia
authors.initials: Z
authors.lastname: Shariat-Madar
authors.forename: Alvin H
authors.initials: AH
authors.lastname: Schmaier
mesh_terms: D000906:Antibodies / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D019921:Biotinylation; D002352:Carrier Proteins; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000378:metabolism*; D005172:Factor XI / Q000378:metabolism*; D015945:Factor XIa / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006801:Humans; D018960:Hyaluronan Receptors; D007633:Keratins / Q000276:immunology; D007700:Kinetics; D019679:Kininogen, High-Molecular-Weight / Q000378:metabolism; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; D015539:Platelet Activation; D011288:Prekallikrein / Q000378:metabolism* / Q000494:pharmacology; D011951:Receptors, Complement / Q000276:immunology; D018000:Receptors, Peptide / Q000276:immunology; D014471:Umbilical Veins; D014568:Urokinase-Type Plasminogen Activator / Q000276:immunology; D015032:Zinc / Q000494:pharmacology
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; C087299:C1QBP protein, human; D002352:Carrier Proteins; D005456:Fluorescent Dyes; D018960:Hyaluronan Receptors; D019679:Kininogen, High-Molecular-Weight; D008562:Membrane Glycoproteins; D024101:Mitochondrial Proteins; D011951:Receptors, Complement; D018000:Receptors, Peptide; C032402:complement 1q receptor; C044865:kininogen receptor; D007633:Keratins; D005172:Factor XI; D011288:Prekallikrein; D015945:Factor XIa; D014568:Urokinase-Type Plasminogen Activator; D016650:Fluorescein-5-isothiocyanate; D015032:Zinc