Decay accelerating activity of complement receptor type 1 (CD35). Two active sites are required for dissociating C5 convertases.
The goal of this study was to identify the site(s) in CR1 that mediate the dissociation of the C3 and C5 convertases. To that end, truncated derivatives of CR1 whose extracellular part is composed of 30 tandem repeating modules, termed complement control protein repeats (CCPs), were generated. Site 1 (CCPs 1-3) alone mediated the decay acceleration of the classical and alternative pathway C3 convertases. Site 2 (CCPs 8-10 or the nearly identical CCPs 15-17) had one-fifth the activity of site 1. In contrast, for the C5 convertase, site 1 had only 0.5% of the decay accelerating activity, while site 2 had no detectable activity. Efficient C5 decay accelerating activity was detected in recombinants that carried both site 1 and site 2. The activity was reduced if the intervening repeats between site 1 and site 2 were deleted. The results indicate that, for the C5 convertases, decay accelerating activity is mediated primarily by site 1. A properly spaced site 2 has an important auxiliary role, which may involve its C3b binding capacity. Moreover, using homologous substitution mutagenesis, residues important in site 1 for dissociating activity were identified. Based on these results, we generated proteins one-fourth the size of CR1 but with enhanced decay accelerating activity for the C3 convertases.
Krych-Goldberg
M
M
Division of Rheumatology, Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Hauhart
R E
RE
Subramanian
V B
VB
Yurcisin
B M
BM
Crimmins
D L
DL
Hourcade
D E
DE
Atkinson
J P
JP
D000595:Amino Acid Sequence; D001665:Binding Sites; D003167:Complement Activation / Q000502:physiology*; D050577:Complement C3-C5 Convertases / Q000378:metabolism*; D003170:Complement Pathway, Alternative / Q000502:physiology; D003171:Complement Pathway, Classical / Q000502:physiology; D017124:Conserved Sequence; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D017463:Receptors, Complement 3b / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid
D010446:Peptide Fragments; D017463:Receptors, Complement 3b; D011994:Recombinant Proteins; D050577:Complement C3-C5 Convertases