A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä
Site-by-site tracking of signal transduction in an azidophenylalanine-labeled bacteriophytochrome with step-scan FTIR spectroscopy (2021)

Kurttila, M., Stucki-Buchli, B., Rumfeldt, J., Schroeder, L., Häkkänen, H., Liukkonen, A., Takala, H., Kottke, T., & Ihalainen, J. (2021). Site-by-site tracking of signal transduction in an azidophenylalanine-labeled bacteriophytochrome with step-scan FTIR spectroscopy. Physical Chemistry Chemical Physics, 23(9), 5615-5628. https://doi.org/10.1039/d0cp06553f

JYU-tekijät tai -toimittajat

Kurttila, Moona
Stucki-Buchli, Brigitte
Rumfeldt, Jessica
Häkkänen, Heikki
Liukkonen, Alli
Takala, Heikki
Ihalainen, Janne

Julkaisun tiedot

Julkaisun kaikki tekijät tai toimittajat: Kurttila, Moona; Stucki-Buchli, Brigitte; Rumfeldt, Jessica; Schroeder, Lea; Häkkänen, Heikki; Liukkonen, Alli; Takala, Heikki; Kottke, Tilman; Ihalainen, Janne

Lehti tai sarja: Physical Chemistry Chemical Physics

ISSN: 1463-9076

eISSN: 1463-9084

Julkaisuvuosi: 2021

Volyymi: 23

Lehden numero: 9

Artikkelin sivunumerot: 5615-5628

Kustantaja: Royal Society of Chemistry (RSC)

Julkaisumaa: Britannia

Julkaisun kieli: englanti

DOI: https://doi.org/10.1039/d0cp06553f

Julkaisun avoin saatavuus: Avoimesti saatavilla

Julkaisukanavan avoin saatavuus: Osittain avoin julkaisukanava

Julkaisu on rinnakkaistallennettu (JYX): https://jyx.jyu.fi/handle/123456789/74502

Tiivistelmä

Signal propagation in photosensory proteins is a complex and multidimensional event. Unraveling such mechanisms site-specifically in real time is an eligible but a challenging goal. Here, we elucidate the site-specific events in a red-light sensing phytochrome using the unnatural amino acid azidophenylalanine, vibrationally distinguishable from all other protein signals. In canonical phytochromes, signal transduction starts with isomerization of an excited bilin chromophore, initiating a multitude of processes in the photosensory unit of the protein, which eventually control the biochemical activity of the output domain, nanometers away from the chromophore. By implementing the label in prime protein locations and running two-color step-scan FTIR spectroscopy on the Deinococcus radiodurans bacteriophytochrome, we track the signal propagation at three specific sites in the photosensory unit. We show that a structurally switchable hairpin extension, a so-called tongue region, responds to the photoconversion already in microseconds and finalizes its structural changes concomitant with the chromophore, in milliseconds. In contrast, kinetics from the other two label positions indicate that the site-specific changes deviate from the chromophore actions, even though the labels locate in the chromophore vicinity. Several other sites for labeling resulted in impaired photoswitching, low structural stability, or no changes in the difference spectrum, which provides additional information on the inner dynamics of the photosensory unit. Our work enlightens the multidimensionality of the structural changes of proteins under action. The study also shows that the signaling mechanism of phytochromes is accessible in a time-resolved and site-specific approach by azido probes and demonstrates challenges in using these labels.

YSO-asiasanat: valokemia; fotobiologia; proteiinit; spektroskopia

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