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Spectroscopic study of the transmembrane domain of a rhodopsin-phosphodiesterase fusion protein from a unicellular eukaryote.

J. Biol. Chem.. 2019; 
WatariMasahito,IkutaTatsuya,YamadaDaichi,ShihoyaWataru,YoshidaKazuho,TsunodaSatoshi P,NurekiOsamu,KandoriHi
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Gene Synthesis The Rh- PDE(full) gene (NCBI Gene ID: 16078606) was synthesized after optimization of a human codon (GenScript) as described previously (15). Get A Quote

摘要

The choanoflagellate contains a chimeric rhodopsin protein composed of an N-terminal rhodopsin (Rh) domain and a C-terminal cyclic nucleotide phosphodiesterase (PDE) domain. The Rh-PDE enzyme light-dependently decreases the concentrations of cyclic nucleotides such as cGMP and cAMP. Photoexcitation of purified full-length Rh-PDE yields an "M" intermediate with a deprotonated Schiff base, and its recovery is much faster than that of the enzyme domain. To gain structural and mechanistic insights into the Rh domain, here we expressed and purified the transmembrane domain of Rh-PDE, Rh-PDE(TMD), and analyzed it with transient absorption, light-induced difference UV-visible, and FTIR spectroscopy method... More

关键词

chromophore,light-activated enzyme,microbial rhodopsin,optogenetics,phosphodiesterase,photoisomerization,photoreceptor,proton transfer,proton transport,rhodopsin,spectroscopy,structural change,structure-func