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Plant and Cell Physiology Advance Access originally published online on August 3, 2006
Plant and Cell Physiology 2006 47(9):1251-1261; doi:10.1093/pcp/pcj095
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© The Author 2006. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Characterization of Cyanobacteriochrome TePixJ from a Thermophilic Cyanobacterium Thermosynechococcus elongatus Strain BP-1

Takami Ishizuka1, Takashi Shimada2, Koji Okajima1, Shizue Yoshihara3, Yuriko Ochiai1, Mitsunori Katayama1 and Masahiko Ikeuchi1,*

1 Department of Life Sciences (Biology), The University of Tokyo, Komaba 3-8-1, Meguro, Tokyo, 153-8902 Japan
2 Tsukuba Proteomics Research Center, SHIMADZU BIOTECH, 3-17-1, Azuma, Tsukuba, 305-0031 Japan
3 Department of Biological Science, Graduate School of Science, Osaka Prefecture University, 1-1 Gakuen-cho, Sakai, Osaka, 599-8531 Japan

* Corresponding author: E-mail, mikeuchi{at}bio.c.u-tokyo.ac.jp; Fax, +81-3-5454-4337.

A putative photoreceptor gene, TepixJ, of a thermophilic cyanobacterium is homologous to SypixJ1 that mediates positive phototaxis in the unicellular motile cyanobacterium Synechocystis sp. PCC 6803. The putative chromophore-binding GAF domain of TePixJ protein was overexpressed as a fusion with a polyhistidine tag (His-TePixJ_GAF) in Synechocystis cells and isolated to homogeneity. The photoreversible conversion of His-TePixJ_GAF showed peaks at 531, 341 and 266 nm for the green light-absorbing form (Pg form), and peaks at 433 and 287 nm for the blue light-absorbing form (Pb form). At 77K, the Pg form fluoresced at 580 nm, while the Pb form did not emit any fluorescence. Mass spectrometry of the tryptic chromopeptide demonstrated that a phycocyanobilin isomer binds to the conserved cysteine at ring A via a thioether bond. It is established that TePixJ and SyPixJ1 are novel photoreceptors in cyanobacteria (‘cyanobacteriochromes’) that are similar, but distinct from the phytochromes and bacteriophytochromes.

(Received May 6, 2006; Accepted July 25, 2006)
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