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each residue type were deuterated. The Val45 side chain forms Bicoid homeodomain bound to the consensus TAATCC DNA-
part of the hydrophobic core of the Bicoid homeodomain, with binding site. J. Mol. Biol. 356, 11371151.
one methyl group being in close contact with the side chains of
Leu40 and Phe8, and the other methyl group in close contact
with Leu31, Ser35, and the highly conserved residue Leu16. In
this case, it is somewhat surprising to observe signicant
conformational heterogeneity for Val45, at least in terms of
signicant, multiple rotamer populations. The authors do not
provide a structural interpretation of their data and acknowl-
edge that further work is necessary to support a conclusion that
conformational heterogeneity contributes in a signicant way to
the complexity of the FTIR signals for Val45 and Lys50.
Perhaps an alternate possibility to explain the Val45 spectral
complexity might be some uctuation of other residues in the
vicinity of the Val45 methyl groups, such as Leu31, Ser35, and
Leu40. One wonders whether data interpretation for Val45
would have been simplied via the use of stereospecically
labeled Val, to eliminate the question of whether slightly
dierent chemical environments of the two methyl groups
contributed to the complexity of the FTIR spectrum.
In summary, this work demonstrates the value of FTIR
spectroscopy in structural studies of proteinDNA complexes,
to complement more commonly used biophysical tools.
Conformational heterogeneity can often manifest itself as
missing electron density in X-ray crystallographic studies, and
in NMR approaches, such heterogeneity can lead to exchange-
broadened resonances or motionally averaged results that
require more extensive studies to unravel. In contrast, the
inherently high temporal and spatial resolution of FTIR
measurements provides the potential for discovery of
conformational heterogeneity on the fastest time scales, thus
allowing for important insights into the molecular mechanisms
of proteinDNA recognition.
AUTHOR INFORMATION
Corresponding Author
*E-mail: mark.rance@uc.edu.
ORCID
Mark Rance: 0000-0003-0664-6024
Funding
M.R. acknowledges support from the National Institute of
General Medical Sciences of the National Institutes of Health
(R01 GM063855).
Notes
The author declares no competing nancial interest.
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