Aromatic
amino acids often flank the transmembrane alpha helices
of integral membrane proteins. By favoring locations within the membrane–water
interface of the lipid bilayer, aromatic residues Trp, Tyr, and sometimes
Phe may serve as anchors to help stabilize a transmembrane orientation.
In this work, we compare the influence of interfacial Trp, Tyr, or
Phe residues upon the properties of tilted helical transmembrane peptides.
For such comparisons, it has been critical to start with no more than
one interfacial aromatic residue near each end of a transmembrane
helix, for example, that of GWALP23 (acetyl-GGALW5(LA)6LW19LAGA-[ethanol]amide). To this end, we have
employed 2H-labeled alanines and solid-state NMR spectroscopy
to investigate the consequences of moving or replacing W5 or W19 in
GWALP23 with selected Tyr, Phe, or Trp residues at the same or proximate
locations. We find that GWALP23 peptides having F5, Y5, or W5 exhibit
essentially the same average tilt and similar dynamics in bilayer
membranes of 1,2-dilauroylphosphatidylcholine (DLPC) or 1,2-dioleoylphosphatidylcholine
(DOPC). When double Tyr anchors are present, in Y4,5GWALP23
the NMR observables are markedly more subject to dynamic averaging
and at the same time are less responsive to the bilayer thickness.
Decreased dynamics are nevertheless observed when ring hydrogen bonding
is removed, such that F4,5GWALP23 exhibits a similar extent
of low dynamic averaging as GWALP23 itself. When F5 is the sole aromatic
group in the N-interfacial region, the dynamic averaging is (only)
slightly more extensive than with W5, Y5, or Y4 alone or with F4,5,
yet it is much less than that observed for Y4,5GWALP23.
Interestingly, moving Y5 to Y4 or W19 to W18, while retaining only
one hydrogen-bond-capable aromatic ring at each interface, maintains
the low level of dynamic averaging but alters the helix azimuthal
rotation. The rotation change is about 40° for Y4 regardless
of whether the host lipid bilayer is DLPC or DOPC. The rotational
change (Δρ) is more dramatic and more complex when W19
is moved to W18, as Δρ is about +90° in DLPC but
about −60° in DOPC. Possible reasons for this curious
lipid-dependent helix rotation could include not only the separation
distances between flanking aromatic or hydrophobic residues but also
the absolute location of the W19 indole ring. For the more usual cases,
when the helix azimuthal rotation shows little dependence on the host
bilayer identity, excepting W18GWALP23, the transmembrane
helices adapt to different lipids primarily by changing the magnitude
of their tilt. We conclude that, in the absence of other functional
groups, interfacial aromatic residues determine the preferred orientations
and dynamics of membrane-spanning peptides. The results furthermore
suggest possibilities for rotational and dynamic control of membrane
protein function.