Chemical characterization and location of ionic interactions involved in the assembly of the C1 complex of human complement.
The C1 complex of human complement comprises two loosely interacting subunits, C1q and the Ca(2+)-dependent C1s-C1r-C1r-C1s tetramer. With a view to gain information on the nature of the ionic interactions involved in C1 assembly, we have studied the effects of the chemical modifications of charged residues of C1q or the tetramer on their ability to reconstitute the C1 complex. Treatment of C1q with pyridoxal-5'-phosphate, acetic anhydride, and citraconic anhydride, as well as with cyclohexanedione and diethylpyrocarbonate, inhibited its ability to associate with C1s-C1r-C1r-C1s. Treatment of the collagen-like fragments of C1q with the same reagents yielded the same effects. Treatment of C1s-C1r-C1r-C1s with 1-ethyl-3-[-3-(dimethylamino) propyl] carbodiimide also prevented C1 assembly, through modification of acidic amino acids which were shown to be located in C1r. Further studies on the location of the interaction sites within C1q, using ligand-blotting and competition experiments with synthetic peptides, were unsuccessful, suggesting that these sites are contributed to by two or three of the C1q chains. It is concluded that C1 assembly involves interactions between acidic amino acids of C1r and lysine (hydroxylysine) and arginine residues located within the collagen-like region of C1q. Sequence comparison with mannan binding protein, another collagen-like molecule which binds the C1s-C1r-C1r-C1s tetramer, suggests Arg A38, and HyL B32, B65, and C29 of C1q as possible interaction sites.
Illy
C
C
Laboratoire d'Enzymologie Moléculaire, Institut de Biologie Structurale, Grenoble, France.
Thielens
N M
NM
Arlaud
G J
GJ
D000089:Acetic Anhydrides / Q000494:pharmacology; D000595:Amino Acid Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry; D002949:Citraconic Anhydrides / Q000494:pharmacology; D037222:Collectins; D003172:Complement C1 / Q000096:biosynthesis* / Q000737:chemistry*; D015922:Complement C1q / Q000737:chemistry; D015923:Complement C1r / Q000737:chemistry / Q000378:metabolism; D003173:Complement C1s / Q000737:chemistry / Q000378:metabolism; D017124:Conserved Sequence; D003514:Cyclohexylamines / Q000494:pharmacology; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D006801:Humans; D007202:Indicators and Reagents; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D011732:Pyridoxal Phosphate / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid
D000089:Acetic Anhydrides; D002352:Carrier Proteins; D002949:Citraconic Anhydrides; D037222:Collectins; D003172:Complement C1; D003514:Cyclohexylamines; D007202:Indicators and Reagents; D046911:Macromolecular Substances; C031800:acetic anhydride; D011732:Pyridoxal Phosphate; D015922:Complement C1q; C030435:1,2-cyclohexanediamine; D015923:Complement C1r; D003173:Complement C1s; D004047:Diethyl Pyrocarbonate