title: Histidine-rich glycoprotein binds fibrin(ogen) with high affinity and competes with thrombin for binding to the gamma'-chain.
abstract: Histidine-rich glycoprotein (HRG) is an abundant protein that binds fibrinogen and other plasma proteins in a Zn(2+)-dependent fashion but whose function is unclear. HRG has antimicrobial activity, and its incorporation into fibrin clots facilitates bacterial entrapment and killing and promotes inflammation. Although these findings suggest that HRG contributes to innate immunity and inflammation, little is known about the HRG-fibrin(ogen) interaction. By immunoassay, HRG-fibrinogen complexes were detected in Zn(2+)-supplemented human plasma, a finding consistent with a high affinity interaction. Surface plasmon resonance determinations support this concept and show that in the presence of Zn(2+), HRG binds the predominant γ(A)/γ(A)-fibrinogen and the γ-chain elongated isoform, γ(A)/γ'-fibrinogen, with K(d) values of 9 nm. Likewise, (125)I-labeled HRG binds γ(A)/γ(A)- or γ(A)/γ'-fibrin clots with similar K(d) values when Zn(2+) is present. There are multiple HRG binding sites on fibrin(ogen) because HRG binds immobilized fibrinogen fragment D or E and γ'-peptide, an analog of the COOH terminus of the γ'-chain that mediates the high affinity interaction of thrombin with γ(A)/γ'-fibrin. Thrombin competes with HRG for γ'-peptide binding and displaces (125)I-HRG from γ(A)/γ'-fibrin clots and vice versa. Taken together, these data suggest that (a) HRG circulates in complex with fibrinogen and that the complex persists upon fibrin formation, and (b) by competing with thrombin for γ(A)/γ'-fibrin binding, HRG may modulate coagulation. Therefore, the HRG-fibrin interaction may provide a novel link between coagulation, innate immunity, and inflammation.
authors.affiliation: Thrombosis and Atherosclerosis Research Institute, McMaster University, Hamilton, Ontario L8L 2X2, Canada; the Departments of Medical Sciences, McMaster University, Hamilton, Ontario L8L 2X2, Canada.
authors.forename: Trang T
authors.initials: TT
authors.lastname: Vu
authors.affiliation: Thrombosis and Atherosclerosis Research Institute, McMaster University, Hamilton, Ontario L8L 2X2, Canada; Medicine, McMaster University, Hamilton, Ontario L8L 2X2, Canada.
authors.forename: Alan R
authors.initials: AR
authors.lastname: Stafford
authors.affiliation: Thrombosis and Atherosclerosis Research Institute, McMaster University, Hamilton, Ontario L8L 2X2, Canada; Medicine, McMaster University, Hamilton, Ontario L8L 2X2, Canada.
authors.forename: Beverly A
authors.initials: BA
authors.lastname: Leslie
authors.affiliation: Thrombosis and Atherosclerosis Research Institute, McMaster University, Hamilton, Ontario L8L 2X2, Canada; Medicine, McMaster University, Hamilton, Ontario L8L 2X2, Canada.
authors.forename: Paul Y
authors.initials: PY
authors.lastname: Kim
authors.affiliation: Thrombosis and Atherosclerosis Research Institute, McMaster University, Hamilton, Ontario L8L 2X2, Canada; Medicine, McMaster University, Hamilton, Ontario L8L 2X2, Canada.
authors.forename: James C
authors.initials: JC
authors.lastname: Fredenburgh
authors.affiliation: Thrombosis and Atherosclerosis Research Institute, McMaster University, Hamilton, Ontario L8L 2X2, Canada; the Departments of Medical Sciences, McMaster University, Hamilton, Ontario L8L 2X2, Canada; Medicine, McMaster University, Hamilton, Ontario L8L 2X2, Canada. Electronic address: weitzj@taari.ca.
authors.forename: Jeffrey I
authors.initials: JI
authors.lastname: Weitz
mesh_terms: D001665:Binding Sites; D004305:Dose-Response Relationship, Drug; D005337:Fibrin / Q000737:chemistry; D005340:Fibrinogen / Q000737:chemistry / Q000378:metabolism*; D015241:Fibrinogens, Abnormal / Q000737:chemistry*; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007074:Immunoglobulin G / Q000737:chemistry; D007700:Kinetics; D008024:Ligands; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D020349:Surface Plasmon Resonance; D013917:Thrombin / Q000737:chemistry*; D015032:Zinc / Q000737:chemistry; D000099053:Histidine-Rich Glycoprotein
chemical_list: D015241:Fibrinogens, Abnormal; D007074:Immunoglobulin G; D008024:Ligands; D010455:Peptides; D011506:Proteins; C527521:fibrinogen gamma'; D000099053:Histidine-Rich Glycoprotein; D005337:Fibrin; D005340:Fibrinogen; D013917:Thrombin; D015032:Zinc