Rhodopsin-cyclases for photocontrol of cGMP/cAMP and 2.3 Å structure of the adenylyl cyclase domain

• Published in: Nature communications Vol. 9; no. 1; pp. 2046 - 15
• Main Authors: Scheib, Ulrike, Broser, Matthias, Constantin, Oana M., Yang, Shang, Gao, Shiqiang, Mukherjee, Shatanik, Stehfest, Katja, Nagel, Georg, Gee, Christine E., Hegemann, Peter
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 24.05.2018; Nature Publishing Group; Nature Portfolio
• Subjects: 631/378/340; 631/45/535/1266; 631/45/607; 631/57/2272; 82/16; 82/29; 82/80; 82/83; 9/74; Adenylyl Cyclases - chemistry; Adenylyl Cyclases - genetics; Adenylyl Cyclases - metabolism; Animals; Binding Sites; Blastocladiomycota - chemistry; Blastocladiomycota - enzymology; Blastocladiomycota - genetics; Crystal structure; Crystallization; Cyclic AMP; Cyclic AMP - chemistry; Cyclic AMP - metabolism; Cyclic GMP; Cyclic GMP - chemistry; Cyclic GMP - metabolism; Cyclic nucleotides; Fungal Proteins - chemistry; Fungal Proteins - metabolism; Guanylate cyclase; Guanylate Cyclase - chemistry; Guanylate Cyclase - genetics; Guanylate Cyclase - metabolism; Humanities and Social Sciences; Models, Molecular; multidisciplinary; Mutation; Nucleotides; Protein Binding; Protein Domains; Rats; Rhodopsin; Rhodopsin - chemistry; Rhodopsin - metabolism; Science; Science (multidisciplinary); Second messengers
• DOI: 10.1038/s41467-018-04428-w
• ISSN: 2041-1723

The cyclic nucleotides cAMP and cGMP are important second messengers that orchestrate fundamental cellular responses. Here, we present the characterization of the rhodopsin-guanylyl cyclase from Catenaria anguillulae (CaRhGC), which produces cGMP in response to green light with a light to dark activity ratio >1000. After light excitation the putative signaling state forms with τ = 31 ms and decays with τ = 570 ms. Mutations (up to 6) within the nucleotide binding site generate rhodopsin-adenylyl cyclases (CaRhACs) of which the double mutated YFP-CaRhAC (E497K/C566D) is the most suitable for rapid cAMP production in neurons. Furthermore, the crystal structure of the ligand-bound AC domain (2.25 Å) reveals detailed information about the nucleotide binding mode within this recently discovered class of enzyme rhodopsin. Both YFP-CaRhGC and YFP-CaRhAC are favorable optogenetic tools for non-invasive, cell-selective, and spatio-temporally precise modulation of cAMP/cGMP with light. Cyclic AMP and cGMP orchestrate a variety of cellular responses. Here, authors characterize the cGMP producing rhodopsin-guanylyl cyclase from C. anguillulae and derived adenylyl cyclase by a biochemical and structural approach which demonstrates the usability of these cyclases for optogenetic applications.