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Transmembrane Helix Integrity versus Fraying To Expose Hydrogen Bonds at a Membrane–Water Interface

Afrose, Fahmida, McKay, Matthew J., Mortazavi, Armin, Suresh Kumar, Vasupradha, Greathouse, Denise V., Koeppe, Roger E.
Biochemistry 2018 v.58 no.6 pp. 633-645
alanine, deuterium, geometry, histidine, hydrogen bonding, isotope labeling, landscapes, membrane proteins, models, nuclear magnetic resonance spectroscopy, peptides, stable isotopes
Transmembrane helices dominate the landscape for many membrane proteins. Often flanked by interfacial aromatic residues, these transmembrane helices also contain loops and interhelix segments, which could help in stabilizing a transmembrane orientation. Using ²H nuclear magnetic resonance spectroscopy to monitor bilayer-incorporated model GWALP23 family peptides, we address systematically the issue of helix fraying in relation to the dynamics and orientation of highly similar individual transmembrane helices. We inserted aromatic (Phe, Trp, Tyr, and His) or non-aromatic residues (Ala and Gly) into positions 4 and 5 adjacent to a core transmembrane helix to examine the side-chain dependency of the transmembrane orientation, dynamics, and helix integrity (extent and location of unraveling). Incorporation of [²H]alanine labels enables one to assess the helicity of the core sequence and the peptide termini. For most of the helices, we observed substantial unwinding involving at least three residues at both ends. For the unique case of histidine at positions 4 and 5, an extended N-terminal unwinding was observed up to residue 7. For further investigation of the onset of fraying, we employed A⁴,⁵GWALP23 with ²H labels at residues 4 and 5 and found that the number of terminal residues involved in the unwinding depends on bilayer thicknesses and helps to govern the helix dynamics. The combined results enable us to compare and contrast the extent of fraying for each related helix, as reflected by the deviation of experimental ²H quadrupolar splitting magnitudes of juxta-terminal alanines A3 and A21 from those represented by an ideal helix geometry.