Abstract
Automation can vastly reduce the cost of experimental labor and thus facilitate high experimental throughput, but little off-the-shelf hardware for the automation of illumination experiments is commercially available. Here, we use inexpensive open-source electronics to add programmable illumination capabilities to a multimode microplate reader. We deploy this setup to characterize light-triggered phenomena in three different sensory photoreceptors. First, we study the photoactivation of Arabidopsis thaliana phytochrome B by light of different wavelengths. Second, we investigate the dark-state recovery kinetics of the Synechocystis sp. blue-light sensor Slr1694 at multiple temperatures and imidazole concentrations; while the kinetics of the W91F mutant of Slr1694 are strongly accelerated by imidazole, the wild-type protein is hardly affected. Third, we determine the light response of the Beggiatoa sp. photoactivatable adenylate cyclase bPAC in Chinese hamster ovary cells. bPAC is activated by blue light in dose-dependent manner with a half-maximal intensity of 0.58 mW cm−2; intracellular cAMP spikes generated upon bPAC activation decay with a half time of about 5 minutes after light switch-off. Taken together, we present a setup which is easily assembled and which thus offers a facile approach to conducting illumination experiments at high throughput, reproducibility and fidelity.
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N. C. Rockwell, D. Duanmu, S. S. Martin, C. Bachy, D. C. Price, D. Bhattacharya, A. Z. Worden, J. C. Lagarias, Proc. Natl. Acad. Sci. U. S. A. 2014 111 3871–3876.
A. Möglich, K. Moffat, Photochem. Photobiol. Sci. 2010 9 1286–1300.
P. Hegemann, Annu. Rev. Plant Biol. 2008 59 167–189.
A. Möglich, X. Yang, R. A. Ayers, K. Moffat, Annu. Rev. Plant Biol. 2010 61 21–47.
M. Iseki, S. Matsunaga, A. Murakami, K. Ohno, K. Shiga, K. Yoshida, M. Sugai, T. Takahashi, T. Hori, M. Watanabe, Nature 2002 415 1047–1051.
M. Gomelsky, G. Klug, Trends Biochem. Sci. 2002 27 497–500.
M. Gauden, I. H. van Stokkum, J. M. Key, D. C. Lührs, R. van Grondelle, P. Hegemann, J. T. Kennis, Proc. Natl. Acad. Sci. U. S. A. 2006 103 10895–10900.
N. C. Rockwell, Y. S. Su, J. C. Lagarias, Annu. Rev. Plant Biol. 2006 57 837–858.
J. Hughes, Annu. Rev. Plant Biol. 2013 64 377–402.
K. Deisseroth, G. Feng, A. K. Majewska, G. Miesenböck, A. Ting, M. J. Schnitzer, J. Neurosci. 2006 26 10380–10386.
M.-H. Ryu, O. V. Moskvin, J. Siltberg-Liberles, M. Gomelsky, J. Biol. Chem. 2010 285 41501–41508.
M. Stierl, P. Stumpf, D. Udwari, R. Gueta, R. Hagedorn, A. Losi, W. Gärtner, L. Petereit, M. Efetova, M. Schwarzel, T. G. Oertner, G. Nagel, P. Hegemann, J. Biol. Chem. 2011 286 1181–1188.
R. J. de Marco, A. H. Groneberg, C.-M. Yeh, L. A. Castillo Ramírez, S. Ryu, Front. Neural Circuits 2013 7 82.
M. Efetova, L. Petereit, K. Rosiewicz, G. Overend, F. Haußig, B. T. Hovemann, P. Cabrero, J. A. T. Dow, M. Schwärzel, J. Cell Sci. 2013 126 778–788.
A. Hartmann, R. D. Arroyo-Olarte, K. Imkeller, P. Hegemann, R. Lucius, N. Gupta, J. Biol. Chem. 2013 288 13705–13717.
M. Ni, J. M. Tepperman, P. H. Quail, Cell 1998 95 657–667.
S. Shimizu-Sato, E. Huq, J. M. Tepperman, P. H. Quail, Nat. Biotechnol. 2002 20 1041–1044.
A. Levskaya, O. D. Weiner, W. A. Lim, C. A. Voigt, Nature 2009 461 997–1001.
W. Laan, M. Gauden, S. Yeremenko, R. van Grondelle, J. T. M. Kennis, K. J. Hellingwerf, Biochemistry 2006 45 51–60.
M. T. Alexandre, J. C. Arents, R. van Grondelle, K. J. Hellingwerf, J. T. Kennis, Biochemistry 2007 46 3129–3137.
S. Anderson, V. Dragnea, S. Masuda, J. Ybe, K. Moffat, C. Bauer, Biochemistry 2005 44 7998–8005.
A. Jung, T. Domratcheva, M. Tarutina, Q. Wu, W. H. Ko, R. L. Shoeman, M. Gomelsky, K. H. Gardner, I. Schlichting, Proc. Natl. Acad. Sci. U. S. A. 2005 102 12350–12355.
H. Yuan, S. Anderson, S. Masuda, V. Dragnea, K. Moffat, C. Bauer, Biochemistry 2006 45 12687–12694.
T. Majerus, T. Kottke, W. Laan, K. Hellingwerf, J. Heberle, ChemPhysChem 2007 8 1787–1789.
C. Bonetti, M. Stierl, T. Mathes, I. H. M. van Stokkum, K. M. Mullen, T. A. Cohen-Stuart, R. van Grondelle, P. Hegemann, J. T. M. Kennis, Biochemistry 2009 48 11458–11469.
S. Masuda, K. Hasegawa, T. Ono, Plant Cell Physiol. 2005 46 1894–1901.
F. Wunder, J.-P. Stasch, J. Hütter, C. Alonso-Alija, J. Hüser, E. Lohrmann, Anal. Biochem. 2005 339 104–112.
F. Wunder, A. Rebmann, A. Geerts, B. Kalthof, Mol. Pharmacol. 2008 73 1235–1243.
L. O. Essen, J. Mailliet, J. Hughes, Proc. Natl. Acad. Sci. U. S. A. 2008 105 14709–14714.
T. R. Berkelman, J. C. Lagarias, Anal. Biochem. 1986 156 194–201.
K. Mukougawa, H. Kanamoto, T. Kobayashi, A. Yokota, T. Kohchi, FEBS Lett. 2006 580 1333–1338.
T. Mathes, C. Vogl, J. Stolz, P. Hegemann, J. Mol. Biol. 2009 385 1511–1518.
J. Mehlhorn, H. Steinocher, S. Beck, J. T. M. Kennis, P. Hegemann, T. Mathes, PLoS One 2013 8 e79006.
G. Nagel, D. Ollig, M. Fuhrmann, S. Kateriya, A. M. Musti, E. Bamberg, P. Hegemann, Science 2002 296 2395–2398.
D. R. Hochbaum, Y. Zhao, S. L. Farhi, N. Klapoetke, C. A. Werley, V. Kapoor, P. Zou, J. M. Kralj, D. Maclaurin, N. Smedemark-Margulies, J. L. Saulnier, G. L. Boulting, C. Straub, Y. K. Cho, M. Melkonian, G. K.-S. Wong, D. J. Harrison, V. N. Murthy, B. L. Sabatini, E. S. Boyden, R. E. Campbell, A. E. Cohen, Nat. Methods 2014 11 825–833.
G. M. Avelar, R. I. Schumacher, P. A. Zaini, G. Leonard, T. A. Richards, S. L. Gomes, Curr. Biol. 2014 24 1234–1240.
M.-H. Ryu, I.-H. Kang, M. D. Nelson, T. M. Jensen, A. I. Lyuksyutova, J. Siltberg-Liberles, D. M. Raizen, M. Gomelsky, Proc. Natl. Acad. Sci. U. S. A. 2014 111 10167–10172.
C. F. Gasser, S. Taiber, C.-M. Yeh, C. H. Wittig, P. Hegemann, S. Ryu, F. Wunder, A. Möglich, Proc. Natl. Acad. Sci. U. S. A. 2014 111 8803–8808.
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Richter, F., Scheib, U.S., Mehlhorn, J. et al. Upgrading a microplate reader for photobiology and all-optical experiments. Photochem Photobiol Sci 14, 270–279 (2015). https://doi.org/10.1039/c4pp00361f
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DOI: https://doi.org/10.1039/c4pp00361f