title: Human complement factor H: isolation of cDNA clones and partial cDNA sequence of the 38-kDa tryptic fragment containing the binding site for C3b.
abstract: We isolated cDNA clones coding for the functionally important tryptic N-terminal 38-kDa fragment of human complement control protein factor H using polyclonal and monoclonal antibodies to screen a human liver cDNA library cloned in a bacterial expression vector, PEX-1. By testing the reactivity of antibodies specific for the recombinant proteins produced by individual clones with proteolytic fragments of serum H the exact position of these cDNA clones within H was mapped. One clone, H-19, coding for the 38-kDa fragment of H was sequenced and found to code for 289 amino acids derived from the 38-kDa N-terminal fragment as well as for the first 108 amino acids belonging to the complementary 142-kDa tryptic fragment. The derived protein sequence could be arranged in 6 highly homologous repeats of about 60 amino acids each, the homology between the repeats being determined by the characteristic position of cysteine, proline, glycine, tyrosine and tryptophane residues. The region coding for the epitope recognized by one of our monoclonal antibodies was localized by subcloning restriction fragments of H-19 into the expression plasmid and testing for the expression of this epitope.
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D003179:Complement C3b / Q000378:metabolism; D003180:Complement C3b Inactivator Proteins / Q000235:genetics* / Q000378:metabolism; D017242:Complement Factor H; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D010449:Peptide Mapping; D011994:Recombinant Proteins / Q000235:genetics
chemical_list: C493495:CFH protein, human; D003180:Complement C3b Inactivator Proteins; D011994:Recombinant Proteins; D003179:Complement C3b; D017242:Complement Factor H; D004247:DNA