Thrombin generates previously unidentified C5 products that support the terminal complement activation pathway.
The coagulation and complement pathways simultaneously promote homeostasis in response to injury but cause tissue damage when unregulated. Mechanisms by which they cooperate are poorly understood. To delineate their interactions, we studied the effects of thrombin and C5 convertase on C5 in purified and plasma-based systems, measuring release of the anaphylatoxin C5a, and generation of C5b, the initial component of the lytic membrane attack complex. Thrombin cleaved C5 poorly at R751, yielding minimal C5a and C5b. However, thrombin efficiently cleaved C5 at a newly identified, highly conserved R947 site, generating previously undescribed intermediates C5(T) and C5b(T). Tissue factor-induced clotting of plasma led to proteolysis of C5 at a thrombin-sensitive site corresponding to R947 and not R751. Combined treatment of C5 with thrombin and C5 convertase yielded C5a and C5b(T), the latter forming a C5b(T)-9 membrane attack complex with significantly more lytic activity than with C5b-9. Our findings provide a new paradigm for complement activation, in which thrombin and C5 convertase are invariant partners, enhancing the terminal pathway via the generation of newly uncovered C5 intermediates. Delineating the molecular links between coagulation and complement will provide new therapeutic targets for diseases associated with excess fibrin deposition and complement activation.
Centre for Blood Research, University of British Columbia, Vancouver, BC, Canada.
Michael J
MJ
Krisinger
Verena
V
Goebeler
Zhen
Z
Lu
Scott C
SC
Meixner
Timothy
T
Myles
Edward L G
EL
Pryzdial
Edward M
EM
Conway
D000818:Animals; D001777:Blood Coagulation; D002645:Chickens; D003167:Complement Activation; D050577:Complement C3-C5 Convertases / Q000378:metabolism; D003182:Complement C5 / Q000276:immunology* / Q000378:metabolism; D004912:Erythrocytes / Q000166:cytology / Q000276:immunology; D006461:Hemolysis; D006801:Humans; D059748:Proteolysis; D015398:Signal Transduction; D013917:Thrombin / Q000276:immunology* / Q000378:metabolism; D013925:Thromboplastin / Q000378:metabolism
D003182:Complement C5; D013925:Thromboplastin; D050577:Complement C3-C5 Convertases; D013917:Thrombin