title: Positional independence and additivity of amino acid replacements on helix stability in monomeric peptides.
abstract: The 17-residue peptide acetylAEAAAKEAAAKEAAAKAamide, described as an autonomous folding unit (Marqusee & Baldwin, 1987), has been used to examine the effect of amino acid replacements on helix stability. Alanine residues(s) at positions 4, 9, and 14 in the peptide sequence were replaced either singly or multiply by either serine or methionine residues with solid-phase peptide synthesis. The thermal dependence of the helix/coil transition of each peptide was observed by far-ultraviolet circular dichroism. Within experimental variation, all three single replacements exhibit a common thermal transition, and all three double replacements exhibit a different common thermal transition. These results suggest that replacement of the central alanine residue in the repeat EAAAK located in the N-terminus, in the middle, or in the C-terminus of the peptide helix has the same effect on helix stability. The melting temperature of each thermal transition was estimated by assuming a linear van't Hoff plot and a change in molar ellipticity of 33,500 deg cm2 dmol-1. Such analysis indicates that each replacement of an alanine residue by a serine residue diminishes the melting temperature by 11 +/- 1 degrees C and that each replacement of an alanine residue by a methionine residue diminishes the melting temperature by 6 +/- 1 degrees C. These results suggest that the effect of these replacements on helix stability is additive.
authors.affiliation: Department of Biochemistry, University of Iowa, Iowa City 52242.
authors.forename: G
authors.initials: G
authors.lastname: Merutka
authors.forename: E
authors.initials: E
authors.lastname: Stellwagen
mesh_terms: D000409:Alanine; D000595:Amino Acid Sequence; D002942:Circular Dichroism; D008969:Molecular Sequence Data; D010455:Peptides; D011487:Protein Conformation; D012694:Serine; D013329:Structure-Activity Relationship; D013696:Temperature
chemical_list: D010455:Peptides; D012694:Serine; D000409:Alanine