Revisiting and Redesigning Light-Activated Cyclic-Mononucleotide Phosphodiesterases

• Published in: Journal of molecular biology Vol. 431; no. 17; pp. 3029 - 3045
• Main Authors: Stabel, Robert, Stüven, Birthe, Hansen, Jan Niklas, Körschen, Heinz G., Wachten, Dagmar, Möglich, Andreas
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 09.08.2019
• Subjects: bacteriophytochrome; cyclic mononucleotide; optogenetics; phosphodiesterase; sensory photoreceptor; cyclic mononucleotide; cNMP; PDE; PAC; optogenetics; bacteriophytochrome; cAMP and cGMP; BphP; phosphodiesterase; PCM; CNG; sensory photoreceptor; Cyclic mononucleotide; Signal transduction; Sensory photoreceptor; Optogenetics; Bacteriophytochrome; Phosphodiesterase
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1016/j.jmb.2019.07.011

As diffusible second messengers, cyclic nucleoside monophosphates (cNMPs) relay and amplify molecular signals in myriad cellular pathways. The triggering of downstream physiological responses often requires defined cNMP gradients in time and space, generated through the concerted action of nucleotidyl cyclases and phosphodiesterases (PDEs). In an approach denoted optogenetics, sensory photoreceptors serve as genetically encoded, light-responsive actuators to enable the noninvasive, reversible, and spatiotemporally precise control of manifold cellular processes, including cNMP metabolism. Although nature provides efficient photoactivated nucleotidyl cyclases, light-responsive PDEs are scarce. Through modular recombination of a bacteriophytochrome photosensor and the effector of human PDE2A, we previously generated the light-activated, cNMP-specific PDE LAPD. By pursuing parallel design strategies, we here report a suite of derivative PDEs with enhanced amplitude and reversibility of photoactivation. Opposite to LAPD, far-red light completely reverts prior activation by red light in several PDEs. These improved PDEs thus complement photoactivated nucleotidyl cyclases and extend the sensitivity of optogenetics to red and far-red light. More generally, our study informs future efforts directed at designing bacteriophytochrome photoreceptors. [Display omitted] •Optogenetics enables precise control of intracellular cyclic-mononucleotide levels.•Bacteriophytochrome modules endow phosphodiesterases with red-light sensitivity.•Modular engineering of a suite of enhanced red‐light-activated phosphodiesterases•Red light-activated phosphodiesterases control ion channel flux in mammalian cells.•Light-activated nucleotidyl cyclases and phosphodiesterases expand optogenetics.