R. M. Jain

Solution structure of dehydropeptides: A CD investigation

A CD investigation of eleven dehydropeptides is reported. The compounds investigated include tri-, tetra-, hepta-, and octapeptides and contain one, two, or three dehydro-phenylalanine (deltaPhe) residues. The peptides showed different CD profiles depending on chain length, position, and number of dehydro residues. The CD data very much complemented that provided by nmr studies, confirming the conformational preference for beta-bend structures in small peptides (tripeptides), and 3(10)-helical or alpha-helical structures in longer peptides. The secondary structures were stable in chloroform solution and were denatured by addition of trifluoroacetic acid. Solvent titration experiments performed by measuring CD as a function of solvent composition provided evidence that the order < or > disorder conformational changes occurred as cooperative transitions.

Conformationally restricted peptides: solution conformation of tetra and hepta peptides containing α,β-dehydrophenylalanine residues in alternate positions

Abstract Two model peptides containing dehydrophenylalanine, a tetrapeptide 1 (Ac- Δ z Phe-Pro-Δ z Phe-Ala-OMe) and a heptapeptide 2 (Boc-Gly-Δ z Phe-Val-Δ z Phe-Ala-Δ z Phe-Leu-OMe) have been synthesised and their solution conformations investigated by NMR and circular dichroism techniques. Assignment of amide protons and their involvement in intramolecular hydrogen bonding have been made by solvent and temperature dependence studies. These conformation studies indicate the presence of an incipient 3 10 -helix in tetrapeptide 1 , with two consecutive β-turns and a right handed 3 10 -helix in heptapeptide 2 . The results establish the potential of Δ z Phe residues to favour 3 10 -helical conformations with δ z Phe occupying alternate positions in the peptide. A comparison of solution conformation of analogous peptides containing Aib residue in place of Δ z Phe is also presented. These residues appear to induce similar conformation constraints in small peptides.

Role of two consecutive α,β-dehydrophenylalanines in peptide structure: crystal and molecular structure of Boc-Leu-ΔPhe-ΔPhe-Ala-Phe-NHMe

An Nα‐protected model pentapeptide containing two consecutive ΔPhe residues, Boc‐Leu‐ΔPhe‐ΔPhe‐Ala‐Phe‐NHMe, has been synthesized by solution methods and fully characterized. 1H‐nmr studies provided evidence for the occurrence of a significant population of a conformer having three consecutive, intramolecularly H‐bonded β‐bends in solution. The solid state structure has been determined by x‐ray diffraction methods. The crystals grown from aqueous methanol are orthorhombic, space group P212121, a = 11.503(2), b = 16.554(2), c = 22.107(3) Å, V = 4209(1) Å,3 and Z = 4. The x‐ray data were collected on a CAD4 diffractometer using CuKa radiation (λ = 1.5418 Å). The structure was determined using direct methods and refined by full‐matrix least‐squares procedure. The R factor is 5.3%. The molecule is characterized by a right handed 310‐helical conformation (〈ϕ〉 = −68.2°, 〈ψ〉 = −26.3°), which is made up of two consecutive type III β‐bends and one type I β‐bend. In the solid state the helical molecules are aligned head‐to‐tail, thus forming long rod like structures. A comparison with other peptide structures containing consecutive ΔPhe residues is also provided. The present study confirms that the ‐ΔPhe‐ΔPhe‐sequence can be accommodated in helical structures.

Helix termination and chain reversal: crystal and molecular structure of the alpha, beta-dehydrooctapeptide Boc-Val-DeltaPhe-Phe-Ala-Leu-Ala-DeltaPhe-Leu-OH.

The crystal structure of the dehydro octapeptide Boc-Val-DeltaPhe-Phe-Ala-Leu-Ala-DeltaPhe-Leu-OH has been determined to atomic resolution by X-ray crystallographic methods. The crystals grown by slow evaporation of peptide solution in methanol/water are orthorhombic, space group P2(1)2(1)2(1). The unit cell parameters are a= 8.404(3), b= 25.598(2) and c= 27.946(3) Angstrom, Z=4. The agreement factor is R = 7.58% for 3636 reflections having (vertical bar Fo vertical bar) > or = 3sigma (vertical bar Fo vertical bar). The peptide molecule is characterised by a 3(10)-helix at the N-terminus and a pi-turn at the C-terminus. This conformation is exactly similar to the helix termination features observed in proteins. The pi-turn conformation observed in the octapeptide is in good agreement with the conformational features of pi-turns seen in some proteins. The alphaL-position in the pi-turn of the octapeptide is occupied by DeltaPhe7, which shows that even bulky residues can be accommodated in this position of the pi-turns. In proteins, it is generally seen that alphaL-position is occupied by glycine residue. No intermolecular head-to-tail hydrogen bonds are observed in solid state structure of the octapeptide. A water molecule located in the unit cell of the peptide molecule is mainly used to hold the peptide molecule together in the crystal. The conformation observed for the octapeptide might be useful to understand the helix termination and chain reversal in proteins and to construct helix terminators for denovo protein design.

IR investigation on dehydrophenylalanine containing model peptides in helical conformation deposited on a crystal surface

Fourier transform ir spectra have been recorded for three 3(10)-helical and one alpha-helical pentapeptides containing dehydrophenylalanine, in a thin solid film, in order to find marker bands for various secondary structures encountered in peptides containing dehydroaminoacids. The peptide solutions were deposited and dried as thin film on zinc selenide crystal surface. This convenient sampling method has provided reliable estimates of peptide secondary structure in solid state. Detailed vibrational assignments in the spectral region between 1200-1700 cm(-1) are reported. In this region, peptide amide I, II, and III vibrations occur. Spectra-structure correlation has been presented based on the amide modes. Comparison of the ir spectra with available crystal structure data provides qualitative support for assignments of ir bands to 3(10)-helical structure and alpha-helical structure in dehydrophenylalanine containing pentapeptides. Band frequency assignments for 3(10)-helical conformation are consistent for all three peptides. All the assignments agree closely with the theoretical predictions. The spectral differences between 3(10)-helical peptides and the alpha-helical peptide have been highlighted. These findings demonstrate that a method based on ir spectroscopy can be developed for a useful approximation of three-dimensional structure of dehydropeptides in solid state. Copyright 1999 John Wiley & Sons, Inc.