Structural features and biologic properties of fragments obtained by limited proteolysis of C3.
Limited proteolysis of the third component of human complement (C3) was performed by using trypsin and streptococcal proteinase and the digests were analyzed for biologic activity. Incubation of C3 with trypsin for 1 min yielded a peptide with smooth muscle-contracting activity but no chemotactic activity, whereas the digest obtained after 15 min of incubation had only chemotactic activity. The conversion of muscle contracting to chemotactic activity could be correlated with the time course of trypsin hydrolysis. Hydrolysis of C3 with streptococcal proteinase gave a digest demonstrating only chemotactic activity. Each digest was resolved on a Sephadex G-100 column and the fractions containing biologic activities were characterized. The amino acid composition of the trypsin fragments, despite having different biologic activities, was remarkably similar, although differences in NH2-terminal amino acids were demonstrable. The fragments obtained by digestion of C3 with the streptococcal proteinase had an amino acid composition different from the trypsin fragments and displayed marked heterogeneity of NH2-terminal residues. These results suggest that slight alterations in the primary structure of C3 fragments may yield significant changes in their biologic activities and that the structural requirements for chemotactic activity are not confined to a single peptide species.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002633:Chemotaxis; D002850:Chromatography, Gel; D003176:Complement C3 / Q000032:analysis* / Q000502:physiology; D003165:Complement System Proteins / Q000032:analysis*; D004589:Electrophoresis, Disc; D006801:Humans; D009119:Muscle Contraction; D010446:Peptide Fragments / Q000032:analysis; D010447:Peptide Hydrolases / Q000494:pharmacology*; D013297:Streptococcus pyogenes / Q000201:enzymology; D013329:Structure-Activity Relationship; D014357:Trypsin / Q000494:pharmacology
D000596:Amino Acids; D003176:Complement C3; D010446:Peptide Fragments; D003165:Complement System Proteins; D010447:Peptide Hydrolases; D014357:Trypsin