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bioRxiv · 10.1101/2021.12.13.472511

Slow conformational changes of blue light sensor BLUF proteins in milliseconds

Tokonami, S.·Onose, M.·Nakasone, Y.·Terazima, M.
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Abstract

BLUF (blue light sensor using flavin) proteins consist of flavin-binding BLUF domains and functional domains. Upon blue light excitation, the hydrogen bond network around the flavin chromophore changes, and the absorption spectrum in the visible region exhibits red-shift. Ultimately, the light information received in the BLUF domain is transmitted to the functional region. It has been believed that this red-shift is complete within nanoseconds. Contrary to this commonly accepted scheme, in this study, slow reaction kinetics were discovered in milliseconds ({tau}1- and {tau}2-phase) for all the BLUF proteins examined (AppA, OaPAC, BlrP1, YcgF, PapB, SyPixD, and TePixD). Despite extensive reports on BLUF, this is the first clear observation of the BLUF protein absorption change with the duration in the millisecond time region. From the measurements of some domain-deleted mutants of OaPAC and two chimeric mutants of PixD proteins, it was found that the slower dynamics ({tau}2-phase) are strongly affected by the size and nature of the C-terminal region adjacent to the BLUF domain. Hence, this millisecond reaction is a significant indicator of conformational changes in the C-terminal region, which is essential for the biological functions. On the other hand, the {tau}1-phase commonly exists in all BLUF proteins, including any mutants. The origin of the slow dynamics was studied using site-specific mutants. These results clearly show the importance of Trp in the BLUF domain. Based on this, a reaction scheme for the BLUF reaction is proposed. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=106 SRC="FIGDIR/small/472511v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@16114dorg.highwire.dtl.DTLVardef@ca015borg.highwire.dtl.DTLVardef@1f3943org.highwire.dtl.DTLVardef@1e89c58_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Tokonami, S., Onose, M., Nakasone, Y., Terazima, M.. 2021-12-15. Slow conformational changes of blue light sensor BLUF proteins in milliseconds. https://doi.org/10.1101/2021.12.13.472511

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