Thrombomodulin enhances complement regulation through strong affinity interactions with factor H and C3b-Factor H complex

Heurich-Sevcenco, M. ORCID: https://orcid.org/0000-0003-0105-2702, Preston, R.J.S., O'Donnell, V. B. ORCID: https://orcid.org/0000-0003-4089-8460, Morgan, B. P. ORCID: https://orcid.org/0000-0003-4075-7676 and Collins, P. W. ORCID: https://orcid.org/0000-0002-6410-1324 2016. Thrombomodulin enhances complement regulation through strong affinity interactions with factor H and C3b-Factor H complex. Thrombosis Research 145 , pp. 84-92. 10.1016/j.thromres.2016.07.017

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Abstract

Introduction 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. Objectives 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.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Pharmacy Medicine
Subjects:	R Medicine > R Medicine (General)
Uncontrolled Keywords:	Complement; Thrombomodulin; Hemolytic assay; Surface plasmon resonance
Additional Information:	This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License
Publisher:	Elsevier
ISSN:	0049-3848
Funders:	NISCHR
Date of First Compliant Deposit:	25 October 2016
Date of Acceptance:	29 July 2016
Last Modified:	24 Nov 2024 17:30
URI:	https://orca.cardiff.ac.uk/id/eprint/95135

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