Comparing Interfacial Trp, Interfacial His and pH Dependence for the Anchoring of Tilted Transmembrane Helical Peptides

Abstract

Charged and aromatic amino acid residues, being enriched toward the terminals of membrane-spanning helices in membrane proteins, help to stabilize particular transmembrane orientations. Among them, histidine is aromatic and can be positively charged at low pH. To enable investigations of the underlying protein-lipid interactions, we have examined the effects of single or pairs of interfacial histidine residues using the constructive low-dynamic GWALP23 (acetyl-GG²ALW⁵LALALALALALALW¹⁹LAG²²A-amide) peptide framework by incorporating individual or paired histidines at locations 2, 5, 19, or 22. Analysis of helix orientation by means of solid-state ²H NMR spectra of labeled alanine residues reveals marked differences with H^2,22 compared to W^2,22. Nevertheless, the properties of membrane-spanning H^2,22WALP23 helices show little pH dependence and are similar to those having Gly, Arg, or Lys at positions 2 and 22. The presence of H5 or H19 influences the helix rotational preference but not the tilt magnitude. H5 affects the helical integrity, as residue 7 unwinds from the core helix; yet, once again, the helix orientation and dynamic properties show little sensitivity to pH. The overall results reveal that the detailed properties of transmembrane helices depend upon the precise locations of interfacial histidine residues.

Keywords: histidine; interfacial aromatic ring; solid-state deuterium NMR; transmembrane helix; tryptophan.