title: Zn2+ mediates high affinity binding of heparin to the αC domain of fibrinogen.
abstract: The nonspecific binding of heparin to plasma proteins compromises its anticoagulant activity by reducing the amount of heparin available to bind antithrombin. In addition, interaction of heparin with fibrin promotes formation of a ternary heparin-thrombin-fibrin complex that protects fibrin-bound thrombin from inhibition by the heparin-antithrombin complex. Previous studies have shown that heparin binds the E domain of fibrinogen. The current investigation examines the role of Zn(2+) in this interaction because Zn(2+) is released locally by platelets and both heparin and fibrinogen bind the cation, resulting in greater protection from inhibition by antithrombin. Zn(2+) promotes heparin binding to fibrinogen, as determined by chromatography, fluorescence, and surface plasmon resonance. Compared with intact fibrinogen, there is reduced heparin binding to fragment X, a clottable plasmin degradation product of fibrinogen. A monoclonal antibody directed against a portion of the fibrinogen αC domain removed by plasmin attenuates binding of heparin to fibrinogen and a peptide analog of this region binds heparin in a Zn(2+)-dependent fashion. These results indicate that the αC domain of fibrinogen harbors a Zn(2+)-dependent heparin binding site. As a consequence, heparin-catalyzed inhibition of factor Xa by antithrombin is compromised by fibrinogen to a greater extent when Zn(2+) is present. These results reveal the mechanism by which Zn(2+) augments the capacity of fibrinogen to impair the anticoagulant activity of heparin.
authors.affiliation: From the Thrombosis and Atherosclerosis Research Institute and.
authors.forename: James C
authors.initials: JC
authors.lastname: Fredenburgh
authors.forename: Beverly A
authors.initials: BA
authors.lastname: Leslie
authors.forename: Alan R
authors.initials: AR
authors.lastname: Stafford
authors.forename: Teresa
authors.initials: T
authors.lastname: Lim
authors.forename: Howard H
authors.initials: HH
authors.lastname: Chan
authors.forename: Jeffrey I
authors.initials: JI
authors.lastname: Weitz
mesh_terms: D000595:Amino Acid Sequence; D000991:Antithrombins / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D001667:Binding, Competitive; D015951:Factor Xa / Q000378:metabolism; D005337:Fibrin / Q000378:metabolism; D005340:Fibrinogen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D020349:Surface Plasmon Resonance; D015032:Zinc / Q000378:metabolism*
chemical_list: D000991:Antithrombins; D010455:Peptides; D005337:Fibrin; D005340:Fibrinogen; D006493:Heparin; D015951:Factor Xa; D015032:Zinc
keywords: Coagulation Factors; Fibrin; Fibrinogen; Heparin; Zinc