title: Engineering of a red-light-activated human cAMP/cGMP-specific phosphodiesterase.
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.
authors.affiliation: Biophysikalische Chemie and.
authors.forename: Carlos
authors.initials: C
authors.lastname: Gasser
authors.affiliation: Lead Discovery, Bayer Pharma AG, 42096 Wuppertal, Germany; and.
authors.forename: Sandra
authors.initials: S
authors.lastname: Taiber
authors.affiliation: Developmental Genetics of the Nervous System, Max Planck Institute for Medical Research, 69120Heidelberg, Germany.
authors.forename: Chen-Min
authors.initials: CM
authors.lastname: Yeh
authors.affiliation: Biophysikalische Chemie and.
authors.forename: Charlotte Helene
authors.initials: CH
authors.lastname: Wittig
authors.affiliation: Experimentelle Biophysik, Institut für Biologie, Humboldt-Universität zu Berlin, 10115 Berlin, Germany;
authors.forename: Peter
authors.initials: P
authors.lastname: Hegemann
authors.affiliation: Developmental Genetics of the Nervous System, Max Planck Institute for Medical Research, 69120Heidelberg, Germany.
authors.forename: Soojin
authors.initials: S
authors.lastname: Ryu
authors.affiliation: Lead Discovery, Bayer Pharma AG, 42096 Wuppertal, Germany; and.
authors.forename: Frank
authors.initials: F
authors.lastname: Wunder
authors.affiliation: Biophysikalische Chemie and andreas.moeglich@hu-berlin.de.
authors.forename: Andreas
authors.initials: A
authors.lastname: Möglich
mesh_terms: D000495:Allosteric Site; D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D000242:Cyclic AMP / Q000737:chemistry*; D006152:Cyclic GMP / Q000737:chemistry*; D054678:Cyclic Nucleotide Phosphodiesterases, Type 2 / Q000235:genetics*; D004926:Escherichia coli / Q000378:metabolism; D017930:Genes, Reporter; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D008027:Light; D008958:Models, Molecular; D010727:Phosphoric Diester Hydrolases / Q000737:chemistry*; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D046939:Synechocystis / Q000378:metabolism; D013696:Temperature; D015027:Zebrafish
chemical_list: D011993:Recombinant Fusion Proteins; D000242:Cyclic AMP; D010727:Phosphoric Diester Hydrolases; D054678:Cyclic Nucleotide Phosphodiesterases, Type 2; C517275:PDE2A protein, human; D006152:Cyclic GMP