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| Title | On the Role of a Conserved Tryptophan in the Chromophore Pocket of Cyanobacteriochrome | ||||
| Date | 2024-06-24 | Attachment | , , , , , , , , | ||
On the Role of a Conserved Tryptophan in the Chromophore Pocket of CyanobacteriochromeBlain-Hartung, M; Sass, GJV; Plaickner, J; Katz, S; Hoang, OT; Mroginski, MA; Esser, N; Budisa, N; Forest, KT; Hildebrandt, P Journal of Molecular Biology, 2024, Volume 436, 168227. The cyanobacteriochrome Slr1393 can be photoconverted between a red (Pr) and green absorbing form (Pg). The recently determined crystal structures of both states suggest a major movement of Trp496 from a stacking interaction with ring D of the phycocyanobilin (PCB) chromophore in Pr to a position outside the chromophore pocket in Pg. Here, we investigated the role of this amino acid during photoconversion in solution using engineered protein variants in which Trp496 was substituted by natural and non -natural amino acids. These variants and the native protein were studied by various spectroscopic techniques (UV-vis absorption, fluorescence, IR, NIR and UV resonance Raman) complemented by theoretical approaches. Trp496 is shown to affect the electronic transition of PCB and to be essential for the thermal equilibrium between Pr and an intermediate state O 600 . However, Trp496 is not required to stabilize the tilted orientation of ring D in Pr, and does not play a role in the secondary structure changes of Slr1393 during the Pr/Pg transition. The present results confirm the re -orientation of Trp496 upon Pr ? Pg conversion, but do not provide evidence of a major change in the microenvironment of this residue. Structural models indicate the penetration of water molecules into the chromophore pocket in both Pr and Pg states and thus water-Trp contacts, which can readily account for the subtle spectral changes between Pr and Pg. Thus, we conclude that reorientation of Trp496 during the Pr-to-Pg photoconversion in solution is not associated with a major change in the dielectric environment in the two states. (c) 2023 Elsevier Ltd. All rights reserved. |
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