Unveiling the nuclear motions of photoreceptor proteins in action is a crucial goal in protein science in order to understand their elaborate mechanisms and how they achieve optimal selectivity and efficiency. Previous studies have provided detailed information on the structures of intermediates that appear during the later stages (>ns) of such photoreception cycles, yet the initial events immediately after photoabsorption remain unclear because of experimental challenges in monitoring nuclear rearrangements on ultrafast timescales, including protein-specific low-frequency motions. Using time-domain Raman probing with sub-7-fs pulses, we obtain snapshot vibrational spectra of photoactive yellow protein and a mutant with high sensitivity, providing insights into the key responses that drive photoreception. Our data show a drastic intensity drop of the excited-state marker band at 135 cm−1 within a few hundred femtoseconds, suggesting a rapid weakening of the hydrogen bond that anchors the chromophore. We also track formation of the first ground-state intermediate over the first few picoseconds and fully characterize its vibrational structure, revealing a substantially-twisted cis conformation.
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This work was partly supported by JSPS KAKENHI Grant Numbers JP16H04102 to S.T., JP25102003 to H.Ka., JP24247030 to M.K. and JP25104005 to T.T. The computations were performed using Research Center for Computational Science, Okazaki, Japan. H.Ku. was supported by RIKEN Special Postdoctoral Researchers (SPDR) programme.
The authors declare no competing financial interests.
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Kuramochi, H., Takeuchi, S., Yonezawa, K. et al. Probing the early stages of photoreception in photoactive yellow protein with ultrafast time-domain Raman spectroscopy. Nature Chem 9, 660–666 (2017). https://doi.org/10.1038/nchem.2717
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