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Origin of β-Hairpin Stability in Solution: Structural and Thermodynamic Analysis of the Folding of a Model Peptide Supports Hydrophobic Stabilization in Water
journal contribution
posted on 1998-02-24, 00:00 authored by Allister J. Maynard, Gary J. Sharman, Mark S. SearleThe origin of the stability of isolated β-hairpins in aqueous
solution is unclear with contrasting
opinions as to the relative importance of interstrand hydrogen bonding,
hydrophobic interactions, and
conformational preferences, the latter being associated largely with
the turn sequence. We have designed an
unconstrained 16-residue peptide that we show folds autonomously in
water to form a β-hairpin that mimics
the two-stranded anti-parallel β-sheet DNA binding motif of the
met repressor dimer. The designed peptide,
with a type I‘ turn (INGK), is shown by CD and a range of NMR
parameters to be appreciably folded (≈50%
at 303 K) in aqueous solution with the predicted alignment of the
peptide backbone. We show that the folding
transition approximates to a two-state model. The hairpin has a
marked temperature-dependent stability, reaching
a maximum value at 303 K in water with both lower and higher
temperatures destabilizing the folded structure.
Van't Hoff analysis of Hα chemical shifts, reveals that folding
is endothermic and entropy-driven in aqueous
solution with a large negative ΔCp, all of
which are reminiscent of proteins with hydrophobic cores,
pointing
to the hydrophobic effect as the dominant stabilizing interaction in
water. We have examined the conformational
properties of the C-terminal β-strand (residues 9−16) in isolation
and have shown that
3JαN values and
backbone
intra- and inter-residue Hα-NH NOE intensities deviate from those
predicted for a random coil, indicating
that the β-strand has a natural predisposition to adopt an extended
conformation in the absence of secondary
structure interactions. A family of β-hairpin structures
calculated from 200 (distance and torsion angle)
restraints
using molecular dynamics shows that the conformation of the hairpin
mimics closely the DNA binding face
of the met repressor dimer (backbone RMSD between
corresponding β-strands of 1.0 ± 0.2 Å).