title: Crystal structure of a complement factor D mutant expressing enhanced catalytic activity.
abstract: Complement factor D is a serine protease regulated by a novel mechanism that depends on conformational changes rather than cleavage of a zymogen for expression of proteolytic activity. The conformational changes are presumed to be induced by the single natural substrate, C3bB, and to result in reversible reorientation of the catalytic center and of the substrate binding site of factor D, both of which have atypical conformations. Here we report that replacement of Ser94, Thr214, and Ser215 of factor D (chymotrypsinogen numbering has been used for comparison purposes) with the corresponding residues of trypsin, Tyr, Ser, and Trp, is sufficient to induce substantially higher catalytic activity associated with a typical serine protease alignment of the catalytic triad residues His57, Asp102, and Ser195. These results provide a partial structural explanation for the low reactivity of "resting-state" factor D toward synthetic substrates.
authors.affiliation: Department of Medicine, University of Alabama at Birmingham 35294-0006, USA.
authors.forename: S
authors.initials: S
authors.lastname: Kim
authors.forename: S V
authors.initials: SV
authors.lastname: Narayana
authors.forename: J E
authors.initials: JE
authors.lastname: Volanakis
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001224:Aspartic Acid; D001483:Base Sequence; D001665:Binding Sites; D002384:Catalysis; D002417:Cattle; D002919:Chymotrypsinogen / Q000737:chemistry; D006639:Histidine; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012694:Serine; D012697:Serine Endopeptidases / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism*; D013552:Swine
chemical_list: D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D001224:Aspartic Acid; D012694:Serine; D006639:Histidine; D002919:Chymotrypsinogen; D012697:Serine Endopeptidases