title: The serine proteinase chain of human complement component C1s. Cyanogen bromide cleavage and N-terminal sequences of the fragments.
abstract: Human complement component C1s was purified from fresh blood by conventional methods of precipitation and chromatography. The single-chain zymogen form was activated by treatment with C1r. Reduction and carboxymethylation then allowed the light chain and heavy chain to be separated on DEAE-Sepharose CL-6B in 8 M-urea. Liquid-phase sequencing of the light chain determined 50 residues from the N-terminus. CNBr-cleavage fragments of the light chain were separated by high-pressure liquid chromatography on gel-permeation and reverse-phase columns. N-Terminal sequencing of these fragments determined the order of a further 138 residues, giving a total of 188 residues or about 75% of the light chain. Seven of these eight sequences could be readily aligned with the amino acid sequences of other serine proteinases. The typical serine proteinase active-site residues are clearly conserved in C1s, and the specificity-related side chain of the substrate-binding pocket is aspartic acid, as in trypsin, consistent with the proteolytic action of C1s on C4 at an arginine residue. Somewhat surprisingly, when the C1s sequence is compared with that of complement subcomponent C1r, the percentage difference (59%) is approximately the same as that found between the other mammalian serine proteinases (56-71%).
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001665:Binding Sites; D003166:Complement Activating Enzymes / Q000302:isolation & purification; D003167:Complement Activation; D003173:Complement C1s; D003488:Cyanogen Bromide; D010450:Endopeptidases / Q000032:analysis*; D006801:Humans; D010446:Peptide Fragments / Q000032:analysis*; D012697:Serine Endopeptidases
chemical_list: D000596:Amino Acids; D010446:Peptide Fragments; D003166:Complement Activating Enzymes; D010450:Endopeptidases; D012697:Serine Endopeptidases; D003173:Complement C1s; D003488:Cyanogen Bromide