Yusuke Ohnishi, Norifumi Muraki, Daiki Kiyota, Hideo Okumura, Seiki Baba, Yoshiaki Kawano, Takashi Kumasaka, Hideaki Tanaka, Genji Kurisu
Journal of biochemistry 2020 Jun 01Plant-type ferredoxin (Fd) is an electron transfer protein in chloroplast. Redox-dependent structural change of Fd controls its association with and dissociation from Fd-dependent enzymes. Among many X-ray structures of oxidized Fd have been reported so far, very likely a given number of them was partially reduced by strong X-ray. To understand the precise structural change between reduced and oxidized Fd, it is important to know whether the crystals of oxidized Fd may or may not be reduced during the X-ray experiment. We prepared the thin plate-shaped Fd crystals from Chlamydomonas reinhardtii and monitored its absorption spectra during experiment. Absorption spectra of oxidized Fd crystals were clearly changed to that of reduced form in an X-ray dose-dependent manner. In another independent experiment, the X-ray diffraction images obtained from different parts of one single crystal were sorted and merged to form two datasets with low and high X-ray doses. An Fo-Fo map calculated from the two datasets showed that X-ray reduction causes a small displacement of the iron atoms in the [2Fe-2S] cluster. Both our spectroscopic and crystallographic studies confirm X-ray dose-dependent reduction of Fd, and suggest a structural basis for its initial reduction step especially in the core of the cluster. © The Author(s) 2020. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.
Yusuke Ohnishi, Norifumi Muraki, Daiki Kiyota, Hideo Okumura, Seiki Baba, Yoshiaki Kawano, Takashi Kumasaka, Hideaki Tanaka, Genji Kurisu. X-ray dose-dependent structural changes of the [2Fe-2S] ferredoxin from Chlamydomonas reinhardtii. Journal of biochemistry. 2020 Jun 01;167(6):549-555
PMID: 32282907
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