Dissection of functional sites in herpesvirus saimiri complement control protein homolog.
Herpesvirus saimiri is known to encode a homolog of human complement regulators named complement control protein homolog (CCPH). We have previously reported that this virally encoded inhibitor effectively inactivates complement by supporting factor I-mediated inactivation of complement proteins C3b and C4b (termed cofactor activity), as well as by accelerating the irreversible decay of the classical/lectin and alternative pathway C3 convertases (termed decay-accelerating activity). To fine map its functional sites, in the present study, we have generated a homology model of CCPH and performed substitution mutagenesis of its conserved residues. Functional analyses of 24 substitution mutants of CCPH indicated that (i) amino acids R118 and F144 play a critical role in imparting C3b and C4b cofactor activities, (ii) amino acids R35, K142, and K191 are required for efficient decay of the C3 convertases, (iii) positively charged amino acids of the linker regions, which are dubbed to be critical for functioning in other complement regulators, are not crucial for its function, and (iv) S100K and G110D mutations substantially enhance its decay-accelerating activities without affecting the cofactor activities. Overall, our data point out that ionic interactions form a major component of the binding interface between CCPH and its interacting partners.
National Centre for Cell Science, Pune University Campus, Ganeshkhind, Pune, India.
Malik Johid
MJ
Reza
Ashish
A
Kamble
Muzammil
M
Ahmad
Musti V
MV
Krishnasastry
Arvind
A
Sahu
D019943:Amino Acid Substitution; D017244:Complement Factor I / Q000235:genetics* / Q000378:metabolism*; D004252:DNA Mutational Analysis; D006567:Herpesvirus 2, Saimiriine / Q000235:genetics / Q000276:immunology*; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism*
D050505:Mutant Proteins; D014764:Viral Proteins; D017244:Complement Factor I