title: The Grateful Infrared: Sequential Protein Structural Changes Resolved by Infrared Difference Spectroscopy.
abstract: The catalytic activity of proteins is a function of structural changes. Very often these are as minute as protonation changes, hydrogen bonding changes, and amino acid side chain reorientations. To resolve these, a methodology is afforded that not only provides the molecular sensitivity but allows for tracing the sequence of these hierarchical reactions at the same time. This feature article showcases results from time-resolved IR spectroscopy on channelrhodopsin (ChR), light-oxygen-voltage (LOV) domain protein, and cryptochrome (CRY). All three proteins are activated by blue light, but their biological role is drastically different. Channelrhodopsin is a transmembrane retinylidene protein which represents the first light-activated ion channel of its kind and which is involved in primitive vision (phototaxis) of algae. LOV and CRY are flavin-binding proteins acting as photoreceptors in a variety of signal transduction mechanisms in all kingdoms of life. Beyond their biological relevance, these proteins are employed in exciting optogenetic applications. We show here how IR difference absorption resolves crucial structural changes of the protein after photonic activation of the chromophore. Time-resolved techniques are introduced that cover the time range from nanoseconds to minutes along with some technical considerations. Finally, we provide an outlook toward novel experimental approaches that are currently developed in our laboratories or are just in our minds ("Gedankenexperimente"). We believe that some of them have the potential to provide new science.
authors.affiliation: Department of Chemistry, Physical and Biophysical Chemistry, Bielefeld University , Universitätsstraße 25, 33615 Bielefeld, Germany.
authors.forename: Tilman
authors.identifier: 0000-0001-8080-9579
authors.initials: T
authors.lastname: Kottke
authors.forename: Víctor A
authors.initials: VA
authors.lastname: Lórenz-Fonfría
authors.affiliation: Experimental Molecular Biophysics, Freie Universität Berlin , Arnimalle 14, 14195 Berlin, Germany.
authors.forename: Joachim
authors.identifier: 0000-0001-6321-2615
authors.initials: J
authors.lastname: Heberle
mesh_terms: D000075402:Channelrhodopsins / Q000737:chemistry*; D056931:Cryptochromes / Q000737:chemistry*; D006860:Hydrogen Bonding; D015394:Molecular Structure; D013055:Spectrophotometry, Infrared / Q000379:methods*
chemical_list: D000075402:Channelrhodopsins; D056931:Cryptochromes