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Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

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Volume 400, Issue 3


Characterization and engineering of photoactivated adenylyl cyclases

Birthe Stüven / Robert Stabel / Robert Ohlendorf / Julian Beck / Roman Schubert / Andreas MöglichORCID iD: https://orcid.org/0000-0002-7382-2772
  • Corresponding author
  • Lehrstuhl für Biochemie, Universität Bayreuth, D-95447 Bayreuth, Germany
  • Institut für Biologie, Humboldt-Universität zu Berlin, D-10115 Berlin, Germany
  • Research Center for Bio-Macromolecules, Universität Bayreuth, D-95447 Bayreuth, Germany
  • Bayreuth Center for Biochemistry and Molecular Biology, Universität Bayreuth, D-95447 Bayreuth, Germany
  • North-Bavarian NMR Center, Universität Bayreuth, D-95447 Bayreuth, Germany
  • orcid.org/0000-0002-7382-2772
  • Email
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  • De Gruyter OnlineGoogle Scholar
Published Online: 2019-01-09 | DOI: https://doi.org/10.1515/hsz-2018-0375


Cyclic nucleoside monophosphates (cNMP) serve as universal second messengers in signal transduction across prokaryotes and eukaryotes. As signaling often relies on transiently formed microdomains of elevated second messenger concentration, means to precisely perturb the spatiotemporal dynamics of cNMPs are uniquely poised for the interrogation of the underlying physiological processes. Optogenetics appears particularly suited as it affords light-dependent, accurate control in time and space of diverse cellular processes. Several sensory photoreceptors function as photoactivated adenylyl cyclases (PAC) and hence serve as light-regulated actuators for the control of intracellular levels of 3′, 5′-cyclic adenosine monophosphate. To characterize PACs and to refine their properties, we devised a test bed for the facile analysis of these photoreceptors. Cyclase activity is monitored in bacterial cells via expression of a fluorescent reporter, and programmable illumination allows the rapid exploration of multiple lighting regimes. We thus probed two PACs responding to blue and red light, respectively, and observed significant dark activity for both. We next engineered derivatives of the red-light-sensitive PAC with altered responses to light, with one variant, denoted DdPAC, showing enhanced response to light. These PAC variants stand to enrich the optogenetic toolkit and thus facilitate the detailed analysis of cNMP metabolism and signaling.

Keywords: adenylyl cyclase; BLUF; optogenetics; phytochrome; sensory photoreceptor; synthetic biology


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About the article

aBirthe Stüven and Robert Stabel: These authors contributed equally to this work.

bPresent address: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

Received: 2018-09-15

Accepted: 2018-12-07

Published Online: 2019-01-09

Published in Print: 2019-02-25

Citation Information: Biological Chemistry, Volume 400, Issue 3, Pages 429–441, ISSN (Online) 1437-4315, ISSN (Print) 1431-6730, DOI: https://doi.org/10.1515/hsz-2018-0375.

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Robert Stabel, Birthe Stüven, Jan Niklas Hansen, Heinz G. Körschen, Dagmar Wachten, and Andreas Möglich
Journal of Molecular Biology, 2019

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