A designed Zn2+-binding amphiphilic polypeptide: energetic consequences of pi-helicity.
The pi-helix is a secondary structure with 4.4 amino acids per helical turn. Although it was proposed in 1952, no experimental support for its existence was obtained until the mid-1980s. While short peptides are unlikely to assume a marginally stable secondary structure spontaneously, they might do so in the presence of appropriate structural constraints. In this paper, we describe a peptide that is designed to assume a pi-helical conformation when stabilized by cetyltrimethylammonium bromide (CTAB) micelles and Zn(2+). In the designed peptide, lipophilic amino acids are placed such that it would be amphiphilic in the pi-helical, but not in the alpha-helical, conformation. Also, two His residues are incorporated with i, i + 5 spacing, designed to allow binding of Zn(2+) in a pi-helical but not an alpha-helical conformation. The peptide was found to form moderately stable monolayers at the air-water interface, with a collapse pressure that almost doubled when there was Zn(2+) in the subphase. Also, CTAB micelles induced a marked increase in the helicity of the peptide. In 50% TFE, the peptide had a CD spectrum consistent with an alpha-helical structure. The addition of 1 mM Zn(2+) to this solvent caused a saturable decline in ellipticity to approximately half of its original value. The peptide also bound Zn(2+) when it was bound to CTAB micelles, with Zn(2+) again inducing a decrease in ellipticity. The peptide had slightly greater affinity for Zn(2+) in the presence of the CTAB than in a 50% TFE solution (K(d) = 3.1 x 10(-4) M in CTAB and 2.3 x 10(-4) M in TFE). van't Hoff analysis indicated that thermal denaturation of the peptide in 50% TFE containing 1 mM Zn(2+) was associated with both enthalpic and entropic changes that were greater than those in the absence of Zn(2+). These observations are all consistent with the proposal that the peptide assumed a pi-helical conformation in the presence of Zn(2+) and CTAB micelles, and has allowed the stability of this rare conformation to be assessed.
Morgan
D M
DM
Department of Pathology, University of Chicago, 5841 South Maryland Avenue, Chicago, Illinois 60637, USA.
Lynn
D G
DG
Miller-Auer
H
H
Meredith
S C
SC
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001057:Apolipoproteins E / Q000138:chemical synthesis / Q000378:metabolism; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000494:pharmacology; D002942:Circular Dichroism; D004795:Enzyme Stability; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis* / Q000378:metabolism*; D011312:Pressure; D011485:Protein Binding / Q000187:drug effects; D015202:Protein Engineering; D017433:Protein Structure, Secondary / Q000187:drug effects; D013499:Surface Properties; D013816:Thermodynamics; D015032:Zinc / Q000378:metabolism*
D001057:Apolipoproteins E; D002593:Cetrimonium Compounds; D010455:Peptides; D015032:Zinc; D000077286:Cetrimonium