Domain swapping reveals complement control protein modules critical for imparting cofactor and decay-accelerating activities in vaccinia virus complement control protein.
Vaccinia virus encodes a structural and functional homolog of human complement regulators named vaccinia virus complement control protein (VCP). This four-complement control protein domain containing secretory protein is known to inhibit complement activation by supporting the factor I-mediated inactivation of complement proteins, proteolytically cleaved form of C3 (C3b) and proteolytically cleaved form of C4 (C4b) (termed cofactor activity), and by accelerating the irreversible decay of the classical and to a limited extent of the alternative pathway C3 convertases (termed decay-accelerating activity [DAA]). In this study, we have mapped the VCP domains important for its cofactor activity and DAA by swapping its individual domains with those of human decay-accelerating factor (CD55) and membrane cofactor protein (MCP; CD46). Our data indicate the following: 1) swapping of VCP domain 2 or 3, but not 1, with homologous domains of decay-accelerating factor results in loss in its C3b and C4b cofactor activities; 2) swapping of VCP domain 1, but not 2, 3, or 4 with corresponding domains of MCP results in abrogation in its classical pathway DAA; and 3) swapping of VCP domain 1, 2, or 3, but not 4, with homologous MCP domains have marked effect on its alternative pathway DAA. These functional data together with binding studies with C3b and C4b suggest that in VCP, domains 2 and 3 provide binding surface for factor I interaction, whereas domain 1 mediates dissociation of C2a and Bb from the classical and alternative pathway C3 convertases, respectively.
Ahmad
Muzammil
M
National Centre for Cell Science, Pune University Campus, Ganeshkhind, Pune, India.
Raut
Sunil
S
Pyaram
Kalyani
K
Kamble
Ashish
A
Mullick
Jayati
J
Sahu
Arvind
A
D018958:CD55 Antigens / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D003165:Complement System Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D017434:Protein Structure, Tertiary; D020349:Surface Plasmon Resonance; D014616:Vaccinia virus / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D014764:Viral Proteins / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*
D018958:CD55 Antigens; D014764:Viral Proteins; C066459:complement-control protein, Vaccinia virus; D003165:Complement System Proteins