Linke, M; Yang, Y; Zienicke, B; Hammam; MAS; Haimberger, T; Zacarias, A; Inomata, K; Lamparter, T; Heyne, K
BIOPHYSICAL JOURNAL 2013, 105, p.1756-1766;
Photoisomerization of biliverdin (BV) chromophore triggers the photoresponse in native Agp1 bacteriophytochrome. Heterogeneity in phytochrome Pr form was discussed to account for the shape of the absorption profile. Different regions of the absorption profile were investigated by angle balanced polarization resolved femtosecond VIS pump – IR probe spectroscopy. We studied the Pr form of Agp1 with its natural chromophore and with a sterically locked 18Et-BV (locked Agp1). We followed the dynamics and orientations of the carbonyl stretching vibrations of ring D and ring A in their ground and electronically excited states. Photoisomerization of ring D is reflected by strong signals of the ring D carbonyl vibration. In contrast, orientational data on ring A show no rotation of ring A upon photoexcitation. Orientational data allows excluding a ZZZasa geometry and corroborates a non-twisted ZZZssa geometry of the chromophore. We found no proof for heterogeneity, but identified a new electronic transition in the absorption profile at 644 nm (S0→S2). Excitation of the S0→S2 transition will introduce a more complex photodynamics compared with S0→S1 transition. Our approach provides fundamental information on disentanglement of absorption profiles, identification of chromophore structures, and determination of molecular groups involved in the photoisomerization process of photoreceptors.