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  Structure-guided design and functional characterization of an artificial red light–regulated guanylate/adenylate cyclase for optogenetic applications

Etzl, S., Lindner, R., Nelson, M. D., & Winkler, A. (2018). Structure-guided design and functional characterization of an artificial red light–regulated guanylate/adenylate cyclase for optogenetic applications. The Journal of Biological Chemistry, 293(23), 9078-9089. doi:10.1074/jbc.RA118.003069.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0001-738C-0 Version Permalink: http://hdl.handle.net/21.11116/0000-0001-738D-F
Genre: Journal Article
Alternative Title : Structure-guided design and functional characterization of an artificial red light-regulated guanylate/adenylate cyclase for optogenetic applications

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 Creators:
Etzl, Stefan, Author
Lindner, Robert1, Author              
Nelson, Matthew D., Author
Winkler, Andreas, Author
Affiliations:
1Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Max Planck Society, ou_1497700              

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Free keywords: adenylate cyclase (adenylyl cyclase) | guanylate cyclase (guanylyl cyclase) | photoreceptor | optogenetics | hydrogen-deuterium exchange | allosteric regulation | adenylyl cyclase | bacteriophytochrome | light regulation | photosensor
 Abstract: Genetically targeting biological systems to control cellular processes with light is the concept of optogenetics. Despite impressive developments in this field, underlying molecular mechanisms of signal transduction of the employed photoreceptor modules are frequently not sufficiently understood to rationally design new optogenetic tools. Here, we investigate the requirements for functional coupling of red light–sensing phytochromes with non-natural enzymatic effectors by creating a series of constructs featuring the Deinococcus radiodurans bacteriophytochrome linked to a Synechocystis guanylate/adenylate cyclase. Incorporating characteristic structural elements important for cyclase regulation in our designs, we identified several red light–regulated fusions with promising properties. We provide details of one light-activated construct with low dark-state activity and high dynamic range that outperforms previous optogenetic tools in vitro and expands our in vivo toolkit, as demonstrated by manipulation of Caenorhabditis elegans locomotor activity. The full-length crystal structure of this phytochrome-linked cyclase revealed molecular details of photoreceptor–effector coupling, highlighting the importance of the regulatory cyclase element. Analysis of conformational dynamics by hydrogen–deuterium exchange in different functional states enriched our understanding of phytochrome signaling and signal integration by effectors. We found that light-induced conformational changes in the phytochrome destabilize the coiled-coil sensor–effector linker, which releases the cyclase regulatory element from an inhibited conformation, increasing cyclase activity of this artificial system. Future designs of optogenetic functionalities may benefit from our work, indicating that rational considerations for the effector improve the rate of success of initial designs to obtain optogenetic tools with superior properties.

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Language(s): eng - English
 Dates: 2018-04-212018-03-212018-04-252018-04-252018-06-01
 Publication Status: Published in print
 Pages: 20
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1074/jbc.RA118.003069
 Degree: -

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Title: The Journal of Biological Chemistry
  Other : JBC
Source Genre: Journal
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Publ. Info: Baltimore, etc. : American Society for Biochemistry and Molecular Biology [etc.]
Pages: - Volume / Issue: 293 (23) Sequence Number: - Start / End Page: 9078 - 9089 Identifier: ISSN: 0021-9258
CoNE: https://pure.mpg.de/cone/journals/resource/954925410826_1