title: Thrombomodulin enhances complement regulation through strong affinity interactions with factor H and C3b-Factor H complex.
abstract: BACKGROUND
  Coagulation and complement systems are simultaneously activated at sites of tissue injury, leading to thrombin generation and opsonisation with C3b. Thrombomodulin (TM) is a cell-bound regulator of thrombin activation, but can also enhance the regulatory activity of complement factor H (FH), thus accelerating the degradation of C3b into inactive iC3b.
  
  
  OBJECTIVE
  This study sought to determine the biophysical interaction affinities of two recombinant TM analogs with thrombin, FH and C3b in order to analyze their ability to regulate serum complement activity.
  
  
  METHODS
  Surface plasmon resonance (SPR) analysis was used to determine binding affinities of TM analogs with FH and C3b, and compared to thrombin as positive control. The capacity of the two recombinant TM analogs to regulate complement in serum was tested in standard complement hemolytic activity assays.
  
  
  RESULTS
  SPR analysis showed that both TM analogs bind FH and C3b-Factor H with nanomolar and C3b with micromolar affinity; binding affinity for its natural ligand thrombin was several fold higher than for FH. At a physiological relevant concentration, TM inhibits complement hemolytic activity in serum via FH dependent and independent mechanisms.
  
  
  CONCLUSIONS
  TM exhibits significant binding affinity for complement protein FH and C3b-FH complex and its soluble form is capable at physiologically relevant concentrations of inhibiting complement activation in serum.
authors: Division of Infection & Immunity and Systems Immunity Research Institute, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, United Kingdom. Electronic address: heurichm@cardiff.ac.uk.
authors: M
authors: M
authors: Heurich
authors: Department of Clinical Medicine, School of Medicine, Trinity College Dublin, Ireland; National Children's Research Centre, Our Lady's Children's Hospital Crumlin, Dublin 12, Ireland.
authors: R J S
authors: RJ
authors: Preston
authors: Division of Infection & Immunity and Systems Immunity Research Institute, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, United Kingdom.
authors: V B
authors: VB
authors: O'Donnell
authors: Division of Infection & Immunity and Systems Immunity Research Institute, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, United Kingdom.
authors: B P
authors: BP
authors: Morgan
authors: Division of Infection & Immunity and Systems Immunity Research Institute, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, United Kingdom.
authors: P W
authors: PW
authors: Collins
mesh_terms: D003167:Complement Activation / Q000502:physiology*; D017242:Complement Factor H / Q000378:metabolism; D006801:Humans; D011485:Protein Binding; D020349:Surface Plasmon Resonance; D018180:Thrombomodulin / Q000378:metabolism*
chemical_list: C493495:CFH protein, human; D018180:Thrombomodulin; D017242:Complement Factor H
keywords: Complement; Hemolytic assay; Surface plasmon resonance; Thrombomodulin