A monomeric human C4b-binding protein (C4bp) more efficiently inactivates C3b than natural C4bp: participation of C-terminal domains in factor I-cofactor activity.
We designed a cDNA construct encoding an artificial membrane molecule consisting of all 8 short consensus repeats (SCRs) of human monomeric C4b-binding protein (C4bp) followed by DAF's GPI anchor, named mC4bp, and expressed the protein on swine endothelial cells (SEC). At the same level of expression, mC4bp protected host cells as effectively as DAF, the most potent complement (C) regulator on the membrane. This result was unexpected from the reported functional properties of natural multimeric C4bp. Here, we investigated the mechanism whereby mC4bp has potent cell-protective activity. Our results were as follows: (1) mC4bp serves more efficiently as a methylamine-treated C3 (C3ma)-inactivating factor I-cofactor than natural C4bp and as efficiently as MCP as a methylamine-treated (C4ma)-inactivating cofactor by fluid-phase cofactor assay: (2) the potency of C3ma inactivation by mC4bp and factor I is quite high compared to those of other cofactors: (3)blocking studies using mAbs against C4bp suggested that both the 48 kDa N-terminal fragment and the C-terminal domain near the portion responsible for bundle formation participate in the high C3ma-inactivating capacity of mC4bp. Thus, acquiring high C3ma-inactivating capacity secondary to monomeric alteration leads to high C regulatory activity of mC4bp. These results infer that mC4bp differs from C4bp in its potent factor I-cofactor activity and is a good candidate as a safeguard against hyperacute rejection of xenografts.
Mikata
S
S
Department of Immunology, Osaka Medical Center for Cancer and Cardiovascular Diseases, Japan.
Miyagawa
S
S
Fukui
A
A
Murakami
Y
Y
Shirakura
R
R
Matsuda
H
H
Hatanaka
M
M
Matsumoto
M
M
Seya
T
T
Suzuki
K
K
Nagasawa
S
S
D003180:Complement C3b Inactivator Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017244:Complement Factor I / Q000378:metabolism; D003169:Complement Inactivator Proteins; D006023:Glycoproteins; D017261:Glycosylphosphatidylinositols; D006801:Humans; D010446:Peptide Fragments; D011487:Protein Conformation; D011951:Receptors, Complement / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid
D003180:Complement C3b Inactivator Proteins; D003169:Complement Inactivator Proteins; D006023:Glycoproteins; D017261:Glycosylphosphatidylinositols; D010446:Peptide Fragments; D011951:Receptors, Complement; D011994:Recombinant Proteins; D017244:Complement Factor I