Engineering of a red-light–activated human cAMP/cGMP-specific phosphodiesterase
- 2 June 2014
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 111 (24), 8803-8808
- https://doi.org/10.1073/pnas.1321600111
Abstract
Sensory photoreceptors elicit vital physiological adaptations in response to incident light. As light-regulated actuators, photoreceptors underpin optogenetics, which denotes the noninvasive, reversible, and spatiotemporally precise perturbation by light of living cells and organisms. Of particular versatility, naturally occurring photoactivated adenylate cyclases promote the synthesis of the second messenger cAMP under blue light. Here, we have engineered a light-activated phosphodiesterase (LAPD) with complementary light sensitivity and catalytic activity by recombining the photosensor module of Deinococcus radiodurans bacterial phytochrome with the effector module of Homo sapiens phosphodiesterase 2A. Upon red-light absorption, LAPD up-regulates hydrolysis of cAMP and cGMP by up to sixfold, whereas far-red light can be used to down-regulate activity. LAPD also mediates light-activated cAMP and cGMP hydrolysis in eukaryotic cell cultures and in zebrafish embryos; crucially, the biliverdin chromophore of LAPD is available endogenously and does not need to be provided exogenously. LAPD thus establishes a new optogenetic modality that permits light control over diverse cAMP/cGMP-mediated physiological processes. Because red light penetrates tissue more deeply than light of shorter wavelengths, LAPD appears particularly attractive for studies in living organisms.This publication has 45 references indexed in Scilit:
- Optogenetic elevation of endogenous glucocorticoid level in larval zebrafishFrontiers in Neural Circuits, 2013
- The S-Helix Determines the Signal in a Tsr Receptor/Adenylyl Cyclase ReporterPublished by Elsevier BV ,2012
- Bright and stable near-infrared fluorescent protein for in vivo imagingNature Biotechnology, 2011
- Light Modulation of Cellular cAMP by a Small Bacterial Photoactivated Adenylyl Cyclase, bPAC, of the Soil Bacterium BeggiatoaJournal of Biological Chemistry, 2011
- Natural and Engineered Photoactivated Nucleotidyl Cyclases for Optogenetic ApplicationsJournal of Biological Chemistry, 2010
- Mechanism for the allosteric regulation of phosphodiesterase 2A deduced from the X-ray structure of a near full-length constructProceedings of the National Academy of Sciences, 2009
- Structure and Signaling Mechanism of Per-ARNT-Sim DomainsStructure, 2009
- Design and Signaling Mechanism of Light-Regulated Histidine KinasesJournal of Molecular Biology, 2008
- Crystal structure of Pseudomonas aeruginosa bacteriophytochrome: Photoconversion and signal transductionProceedings of the National Academy of Sciences, 2008
- Channelrhodopsin-2, a directly light-gated cation-selective membrane channelProceedings of the National Academy of Sciences, 2003