title: Effects of a heparin-binding protein on blood coagulation and platelet function.
abstract: The objective of this study was to characterize the heparin-binding properties of a protein secreted by mouse myeloma cells. The characterization was performed using clinical assays, such as heparin activity assays and heparin-induced thrombocytopenia (HIT) platelet activation assays. The tests were performed in the presence of heparin, low-molecular-weight heparins (LMWH), or heparinoids and either heparin-binding protein (HBP) or saline to determine whether the HBP affects the activity of heparins. The characterization of the HBP using heparin activity assays showed that the HBP shortened the prolonged clotting times of the activated partial thromboplastin time (aPTT) and thrombin clotting time induced by high concentrations of unfractionated heparin. The chromogenic assays for antithrombin (AT), thrombin inhibition, and factor Xa inhibition demonstrated that this effect is related to heparin concentrations below 0.5 IU/ml. The Heptest assay did not detect these differences. The HBP did not modify the anticoagulant effect of any LMWH or low- or high-sulfated glycosaminoglycans in the aPTT assay. Activation of donor platelets in the presence of unfractionated heparin, platelet factor 4 (PF4), and HIT-serum was not counteracted by the HBP in any of the assays. The characterization of the HBP using a PF4-enzyme-linked immunosorbent assay (ELISA) confirmed the lack of structural identity with PF4. However, the optical density data indicated that the protein structure may be similar to PF4 by binding to a PF4 antibody. These data suggest that the HBP isolated from mouse myeloma cells has a low affinity to heparin and interacts with the secondary binding site to AT and also perhaps to PF4.
authors.affiliation: First Department of Medicine, University Hospital, Mannheim, Germany.
authors.forename: P
authors.initials: P
authors.lastname: Kaiser
authors.forename: J
authors.initials: J
authors.lastname: Harenberg
authors.forename: J M
authors.initials: JM
authors.lastname: Walenga
authors.forename: G
authors.initials: G
authors.lastname: Huhle
authors.forename: C
authors.initials: C
authors.lastname: Giese
authors.forename: M
authors.initials: M
authors.lastname: Prechel
authors.forename: D
authors.initials: D
authors.lastname: Hoppensteadt
authors.forename: J
authors.initials: J
authors.lastname: Fareed
mesh_terms: D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects*; D001780:Blood Coagulation Tests; D002352:Carrier Proteins / Q000494:pharmacology*; D006493:Heparin / Q000378:metabolism*; D051379:Mice; D009101:Multiple Myeloma / Q000473:pathology; D009363:Neoplasm Proteins / Q000378:metabolism* / Q000494:pharmacology*; D015539:Platelet Activation / Q000187:drug effects*; D010979:Platelet Function Tests; D014407:Tumor Cells, Cultured
chemical_list: D002352:Carrier Proteins; D009363:Neoplasm Proteins; D006493:Heparin