Variation in carotenoid-protein interaction in bird feathers produces novel plumage coloration.

Light absorption by carotenoids is known to vary substantially with the shape or conformation of the pigment molecule induced by the molecular environment, but the role of interactions between carotenoid pigments and the proteins to which they are bound, and the resulting impact on organismal coloration, remain unclear. Here, we present a spectroscopic investigation of feathers from the brilliant red scarlet ibis (Eudocimus ruber, Threskiornithidae), the orange-red summer tanager (Piranga rubra, Cardinalidae) and the violet-purple feathers of the white-browed purpletuft (Iodopleura isabellae, Tityridae). Despite their striking differences in colour, all three of these feathers contain canthaxanthin (β,β-carotene-4,4'-dione) as their primary pigment. Reflectance and resonance Raman (rR) spectroscopy were used to investigate the induced molecular structural changes and carotenoid-protein interactions responsible for the different coloration in these plumage samples. The results demonstrate a significant variation between species in the peak frequency of the strong ethylenic vibration (ν(1)) peak in the rR spectra, the most significant of which is found in I. isabellae feathers and is correlated with a red-shift in canthaxanthin absorption that results in violet reflectance. Neither polarizability of the protein environment nor planarization of the molecule upon binding can entirely account for the full extent of the colour shift. Therefore, we suggest that head-to-tail molecular alignment (i.e. J-aggregation) of the protein-bound carotenoid molecules is an additional factor.