Abstract
The protein encoded by the gene slr1393 from the cyanobacterium Synechocystis sp. PCC6803 (Slr1393) is composed of three GAF domains, a PAS domain, and a histidine kinase motif. The third GAF domain (referred to as GAF3) was previously characterized as the sole domain in this protein, being able to carry phycocyanobilin (PCB) as the chromophore and to accomplish photochemistry. GAF3 shows photochromicity, and is able to switch between a red-absorbing parental state (GAF3R, λmax = 649 nm) and a green-absorbing photoproduct state (GAF3G, λmax = 536 nm) upon appropriate irradiation. In this study we have determined the photochemical quantum yields for the interconversion between both forms using two methods: an “absolute” method and a reference-based control. The latter is a comparative procedure which exploits a well-characterized blue-light photoreceptor, YtvA from Bacillus subtilis, and the cyanobacterial phytochrome Cph1 as actinometers. The former is an ad hoc developed, four laser-based setup where two cw lasers provide the pump beams to induce photoswitching (red to green and green to red, respectively) and two cw lasers simultaneously monitor the appearance and disappearance of the two species. Interestingly, fit analysis of the recorded transient absorbance changes provided a quantum yield for the green → red conversion (≈0.3) at least three times larger than for the red → green conversion (≈0.08). These data are in agreement with the results from the comparative method documenting the usefulness of the ‘direct’ method developed here for quantum yields’ determination. The light-induced switching capability of this photochromic protein allowed measuring the kinetics of GAF3 immobilized on a glass plate, and within living, overexpressing Escherichia coli cells.
Similar content being viewed by others
References
M. Ohmori, M. Ikeuchi, N. Sato, P. Wolk, T. Kaneko, T. Ogawa, M. Kanehisa, S. Goto, S. Kawashima, S. Okamoto, H. Yoshimura, H. Katoh, T. Fujisawa, S. Ehira, A. Kamei, S. Yoshihara, R. Narikawa, S. Tabata, DNA Res. 2001 8 271–284.
M. Ikeuchi, T. Ishizuka, Photochem. Photobiol. Sci. 2008 7 1159–1167.
P. Jones, D. Binns, H.-Y. Chang, M. Fraser, W. Li, C. McAnulla, H. McWilliam, J. Maslen, A. Mitchell, G. Nuka, S. Pesseat, A. F. Quinn, A. Sangrador-Vegas, M. Scheremetjew, S.-Y. Yong, R. Lopez, S. Hunter, Bioinformatics 2014 30 1236–1240.
S. Sharda, R. Shah, W. Gärtner, Eur. Biophys. J. 2007 36 815–821.
R. Narikawa, Y. Fukushima, T. Ishizuka, S. Itoh, M. Ikeuchi, J. Mol. Biol. 2008 380 844–855.
T. Ishizuka, T. Shimada, K. Okajima, S. Yoshihara, Y. Ochiai, M. Katayama, M. Ikeuchi, Plant Cell Physiol. 2006 47 1251–1261.
Y. Chen, J. Zhang, J. Luo, J.-M. Tu, X.-L. Zeng, J. Xie, M. Zhou, J.-Q. Zhao, H. Scheer, K.-H. Zhao, FEBS J. 2012 279 40–54.
G. Enomoto, Y. Hirose, R. Narikawa, M. Ikeuchi, Biochemistry 2012 51 3050–3058.
T. Ishizuka, A. Kamiya, H. Suzuki, R. Narikawa, T. Noguchi, T. Kohchi, K. Inomata, M. Ikeuchi, Biochemistry 2011 50 953–961.
E. S. Burgie, Joseph M. Walker, George N. Phillips Jr., Richard D. Vierstra, Structure 2013 21 88–97.
N. C. Rockwell, S. S. Martin, K. Feoktistova, J. C. Lagarias, Proc. Natl. Acad. Sci. U. S. A. 2011 108 11854–11859.
H. Brock, B. P. Ruzsicska, T. Arai, W. Schlamann, A. R. Holzwarth, S. E. Braslavsky, Biochemistry 1987 26 1412–1417.
J. Zhang, X.-J. Wu, Z.-B. Wang, Y. Chen, X. Wang, M. Zhou, H. Scheer, K.-H. Zhao, Angew. Chem., Int. Ed. 2010 49 5456–5458.
A. Losi, W. Gärtner, S. Raffelberg, F. C. Zanacchi, P. Bianchini, A. Diaspro, C. Mandalari, S. Abbruzzetti, C. Viappiani, Photochem. Photobiol. Sci. 2013 12 231–235.
G. Enomoto, R. Nomura, T. Shimada, N.-N. Win, R. Narikawa, M. Ikeuchi, J. Biol. Chem. 2014 289 24801–24809.
D. Tischer, O. D. Weiner, Nat. Rev. Mol. Cell Biol. 2014 15 551–558.
P. W. Kim, L. H. Freer, N. C. Rockwell, S. S. Martin, J. C. Lagarias, D. S. Larsen, Biochemistry 2012 51 619–630.
Y. Fukushima, M. Iwaki, R. Narikawa, M. Ikeuchi, Y. Tomita, S. Itoh, Biochemistry 2011 50 6328–6339.
X.-L. Xu, A. Gutt, J. Mechelke, S. Raffelberg, K. Tang, D. Miao, L. Valle, C. D. Borsarelli, K.-H. Zhao, W. Gärtner, ChemBioChem 2014 15 1190–1199.
A. F. E. Hauck, S. J. O. Hardman, R. J. Kutta, G. M. Greetham, D. J. Heyes, N. S. Scrutton, J. Biol. Chem. 2014 289 17747–17757.
P. W. Kim, L. H. Freer, N. C. Rockwell, S. S. Martin, J. C. Lagarias, D. S. Larsen, J. Am. Chem. Soc. 2012 134 130–133.
R. Narikawa, T. Ishizuka, N. Muraki, T. Shiba, G. Kurisu, M. Ikeuchi, Proc. Natl. Acad. Sci. U. S. A. 2013 110 918–923.
E. S. Burgie, A. N. Bussell, J. M. Walker, K. Dubiel, R. D. Vierstra, Proc. Natl. Acad. Sci. U. S. A. 2014 111 10179–10184.
C. C. Cornilescu, G. Cornilescu, E. S. Burgie, J. L. Markley, A. T. Ulijasz, R. D. Vierstra, J. Biol. Chem. 2014 289 3055–3065.
S. Raffelberg, M. Mansurova, W. Gärtner, A. Losi, J. Am. Chem. Soc. 2011 133 5346–5356.
A. M. Brouwer, Pure Appl. Chem. 2011 83 2213–2228.
J. Mailliet, G. Psakis, K. Feilke, V. Sineshchekov, L.-O. Essen, J. Hughes, J. Mol. Biol. 2011 413 115–127.
V. Voliani, R. Bizzarri, R. Nifosi, S. Abbruzzetti, E. Grandi, C. Viappiani, F. Beltram, J. Phys. Chem. B 2008 112 10714–10722.
J. C. Lagarias, J. M. Kelly, K. L. Cyr, W. O. Smith, Photochem. Photobiol. 1987 46 5–13.
T. Lamparter, F. Mittmann, W. Gärtner, T. Bärner, E. Hartmann, J. Hughes, Proc. Natl. Acad. Sci. U. S. A. 1997 94 11792–11797.
C.-W. Chang, S. M. Gottlieb, P. W. Kim, N. C. Rockwell, J. C. Lagarias, D. S. Larsen, J. Phys. Chem. B 2013 117 11229–11238.
A. Losi, E. Polverini, B. Quest, W. Gärtner, Biophys. J. 2002 82 2627–2634.
S. J. Hagen, W. A. Eaton, J. Chem. Phys. 1996 104 3395–3398.
L. Cordone, G. Cottone, S. Giuffrida, G. Palazzo, G. Venturoli, C. Viappiani, Biochim. Biophys. Acta, Proteins Proteomics 2005 1749 252–281.
F. Pennacchietti, S. Abbruzzetti, A. Losi, C. Mandalari, R. Bedotti, C. Viappiani, F. C. Zanacchi, A. Diaspro, W. Gärtner, PLoS One 2014 9 e107489.
F. Cannone, M. Caccia, S. Bologna, A. Diaspro, G. Chirico, Microsc. Res. Tech. 2004 65 186–193.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Pennacchietti, F., Losi, A., Xu, Xl. et al. Photochromic conversion in a red/green cyanobacteriochrome from Synechocystis PCC6803: quantum yields in solution and photoswitching dynamics in living E. coli cells. Photochem Photobiol Sci 14, 229–237 (2015). https://doi.org/10.1039/c4pp00337c
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1039/c4pp00337c