1H NMR studies of prototypical helical designer peptides. A comparative study of the amide chemical shift dependency on temperature and polypeptide sequence.
A series of designer alpha-helical peptides with hydrophobic residues located at different positions along the sequence (PH-1.0 = LYQELQKLTQTLK, PH-1.19 = LYQELQKLTQTFK, PH-1.12 = LYQELQKLLQTLK, PH-1.13 = LYQELQKLTLTLK, PH-1.4 = LYQELQKLTQTTK) were analyzed using one- and two-dimensional NMR methods (TOCSY and NOESY). The central feature of these designer peptides is the incorporation of a maximal hydrophobic strip which may play a role in antigen processing and the nucleation of alpha-helices in proteins (J. Immunol. 145, 899, 1990). Using the 2D-NMR, sequence specific assignments and NOE connectivities were determined in all peptides when dissolved in H2O/TFE mixtures. NOE connectivities indicated that all these peptides are helical in this medium. An unusually large number of NOEs was found for all these designer peptides. This is in accord with ultracentrifugation studies that showed that PH-1.0 forms a trimer in 50% H2O/TFE mixtures. Other peptides in the series behave in similar manner as PH-1.0. The structural differences among these peptides was addressed using the backbone amide chemical shift temperature coefficients, [symbol: see text], and the differences between the observed and random coil values, delta delta HN. The delta delta HN patterns along the peptide sequence are consistent with those expected for amphiphilic alpha-helices, where most delta delta HN values are below zero. However, no significant differences among the peptides in this series can be detected on the delta delta HN patterns, with the exception of PH-1.12. The [symbol: see text] values reveal differences among the peptides of the series. The patterns of [symbol: see text] along the peptide sequences are similar to that found for delta delta HN for PH-1.0, PH-1.19 and PH-1.4. The other peptides in the series, PH-1.12 and PH-1.13, showed different patterns for [symbol: see text]. The latter parameter was used to evaluate the helicity of this series of peptides. According to this parameter the relative helicity of this series is as follows: PH-1.12 > PH-1.0 > PH-1.4 > PH-1.19 > PH-1.13 The NMR data shown here correlated well with the helical propensities predicted for polypeptide sequences using statistical arguments (Proc. Natl. Acad. Sci. USA, 90, 9100, 1993).
Vinogradov
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Department of Chemistry and Biochemistry, Florida Atlantic University, Boca Raton, USA.
Marí
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Humphreys
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Wright
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D006860:Hydrogen Bonding; D009682:Magnetic Resonance Spectroscopy; D010455:Peptides / Q000737:chemistry*; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D014270:Trifluoroethanol / Q000494:pharmacology
D010455:Peptides; D014270:Trifluoroethanol