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Circular dichroism analysis of cyclic β-helical peptides adsorbed on planar fused quartz.

Research paper by Kenan P KP Fears, Dmitri Y DY Petrovykh, Sara J SJ Photiadis, Thomas D TD Clark

Indexed on: 13 Jul '13Published on: 13 Jul '13Published in: Langmuir



Abstract

Conformational changes of three cyclic β-helical peptides upon adsorption onto planar fused-quartz substrates were detected and analyzed by far-ultraviolet (UV) circular dichroism (CD) spectroscopy. In trifluoroethanol (TFE), hydrophobic peptides, Leu β and Val β, form left- and right-handed helices, respectively, and water-soluble peptide WS β forms a left-handed helix. Upon adsorption, CD spectra showed a mixture of folded and unfolded conformations for Leu β and Val β and predominantly unfolded conformations for WS β. X-ray photoelectron spectroscopy (XPS) provided insight about the molecular mechanisms governing the conformational changes, revealing that ca. 40% of backbone amides in Leu β and Val β were interacting with the hydrophilic substrate, while only ca. 15% of the amines/amides in WS β showed similar interactions. In their folded β-helical conformations, Leu β and Val β present only hydrophobic groups to their surroundings; hydrophilic surface groups can only interact with backbone amides if the peptides change their conformation. Conversely, as a β helix, WS β presents hydrophilic side chains to its surroundings that could, in principle, interact with hydrophilic surface groups, with the peptide retaining its folded structure. Instead, the observed unfolded surface conformation for WS β and the relatively small percentage of surface-bound amides (15 versus 40% for Leu β and Val β) suggest that hydrophilic surface groups induce unfolding. Upon this surface-induced unfolding, WS β interacts with the surface preferentially via hydrophilic side chains rather than backbone amides. In contrast, the unfolded β-hairpin-like form of WS β does not irreversibly adsorb on fused quartz from water, highlighting that solvation effects can be more important than initial conformation in governing peptide adsorption. Both label-free methods demonstrated in this work are, in general, applicable to structural analysis of a broad range of biomolecules adsorbed on transparent planar substrates, the surface properties of which could be customized.