Temperature coefficients of amide proton NMR resonance frequencies in trifluoroethanol: a monitor of intramolecular hydrogen bonds in helical peptides.
2D 1H NMR spectroscopy of two alpha-helical peptides which differ in their amphipathicity has been used to investigate the relationships between amide-proton chemical shifts, amide-proton exchange rates, temperature, and trifluoroethanol (TFE) concentration. In 50% TFE, in which the peptides are maximally helical, the amide-proton chemical shift and temperature coefficient patterns are very similar to each other in each peptide. Temperature coefficients from -10 to -6 ppb/K, usually indicative of the lack of intramolecular hydrogen bonds, were observed even for hydrophobic amino acids in the center of the alpha-helices. However, slow hydrogen isotope exchange for residues from 4 to 16 in both 18-mer helices indicates intact intramolecular hydrogen bonds over most of the length of these peptides. Based on these anomalous observations, we suggest that the pattern of amide-proton shifts in alpha-helices in H20/TFE solvents is dominated by bifurcated intermolecular hydrogen-bond formation between the backbone carbonyl groups and TFE. The amide-proton chemical shift changes with increasing temperature may be interpreted by a disruption of intermolecular hydrogen bonds between carbonyl groups and the TFE in TFE/water rather than by the length of intramolecular hydrogen bonds in alpha-helices.
Rothemund
S
S
Research Institute of Molecular Pharmacology, Berlin, Germany.
Weisshoff
H
H
Beyermann
M
M
Krause
E
E
Bienert
M
M
Mügge
C
C
Sykes
B D
BD
Sönnichsen
F D
FD
D006859:Hydrogen; D009682:Magnetic Resonance Spectroscopy; D010455:Peptides / Q000737:chemistry*; D017510:Protein Folding; D013696:Temperature; D014270:Trifluoroethanol / Q000737:chemistry*
D010455:Peptides; D014270:Trifluoroethanol; D006859:Hydrogen