Conformational analysis of COOH-terminal segments of human C3a. Evidence of ordered conformation in an active 21-residue peptide

Human C3a, a 77-residue fragment released during activation of the serum complement system, is a potent spasmogen that contracts a variety of smooth muscle tissues and enhances vascular permeability. Previous studies have suggested that a 5-residue, COOH-terminal segment of C3a constitutes the essential active site of this molecule; however, the pentapeptide is 1000-fold less active than C3a. Human C3a 57-77, a synthetic segment containing the 21 COOH-terminal residues of C3a, exhibits potencies nearly equivalent to those of natural C3a in several biologic assay systems. The circular dichroism spectra of synthetic peptides corresponding to sequences 57-77, 65-77, and 73-77 in human C3a were measured in water and trifluoroethanol. The CD spectra in the far-UV region indicate that each C3a peptide assumes a random coil conformation in aqueous solution with little evidence of alpha-helical structure. However, C3a peptide 57-77 assumes predominantly an alpha-helical conformation (47%) in 25% trifluoroethanol, while the shorter tridecapeptide 65-77 and pentapeptide 73-77 appear by CD to contain beta-turn conformations only Crystallographic analysis of human C3a indicated that the NH2-terminal portion of peptide 57-77 adopts an alpha-helical structure and that the COOH-terminal portion, including residues 73-77, contains an irregular fold much like a beta-turn. Since C3a peptide 57-77 exhibits activities qualitatively and quantitatively similar to natural C3a, we propose that this synthetic peptide adopts a helical conformation when bound to its cellular receptor which corresponds to that in the intact C3a molecule. Consequently, the NH2-terminal portion (residues 1-21) and the disulfide-linked core region (residues 22-57) in intact C3a serve primarily to stabilize ordered conformation in the COOH-terminal region (residues 58-77) and thereby orient side chains at the essential active site for optimal receptor interaction.