2D-IR study of a photoswitchable isotope-labeled alpha-helix

Title2D-IR study of a photoswitchable isotope-labeled alpha-helix
Publication TypeJournal Article
Year of Publication2010
AuthorsBackus E.HG, Bloem R., Donaldson P.M, Ihalainen J.A, Pfister R., Paoli B., Caflisch A., Hamm P.
JournalThe Journal of Physical Chemistry B
Volume114
Issue10
Pagination3735-3740
Date Published2010 Mar 18
Type of ArticleResearch Article
ISSN1520-5207
KeywordsAmino Acid Sequence, Carbon Isotopes, Circular Dichroism, Hydrogen Bonding, Isotope Labeling, Oxygen Isotopes, Peptides, Protein Folding, Protein Structure, Secondary, Spectrophotometry, Infrared
Abstract

A series of photoswitchable, α-helical peptides were studied using two-dimensional infrared spectroscopy (2D-IR). Single-isotope labeling with 13C18O at various positions in the sequence was employed to spectrally isolate particular backbone positions. We show that a single 13C18O label can give rise to two bands along the diagonal of the 2D-IR spectrum, one of which is from an amide group that is hydrogen-bonded internally, or to a solvent molecule, and the other from a non-hydrogen-bonded amide group. The photoswitch enabled examination of both the folded and unfolded state of the helix. For most sites, unfolding of the peptide caused a shift of intensity from the hydrogen-bonded peak to the non-hydrogen-bonded peak. The relative intensity of the two diagonal peaks gives an indication of the fraction of molecules hydrogen-bonded at a certain location along the sequence. As this fraction varies quite substantially along the helix, we conclude that the helix is not uniformly folded. Furthermore, the shift in hydrogen bonding is much smaller than the change of helicity measured by CD spectroscopy, indicating that non-native hydrogen-bonded or mis-folded loops are formed in the unfolded ensemble.

DOI10.1021/jp911849n
pubindex

0127

Alternate JournalJ. Phys. Chem. B
PubMed ID20166694
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