Journal ArticleDOI
Crystal structure of metarhodopsin II.
Hui-Woog Choe,Yong Ju Kim,Jung Hee Park,Takefumi Morizumi,Takefumi Morizumi,Emil F. Pai,Norbert Krauss,Klaus Peter Hofmann,Klaus Peter Hofmann,Patrick Scheerer,Oliver P. Ernst,Oliver P. Ernst +11 more
TLDR
By comparison with early photoproducts, it is proposed how retinal translocation and rotation induce the gross conformational changes characteristic for Meta’II, and the structures can now serve as models for the large GPCR family.Abstract:
Structural studies of active states of the visual pigment rhodopsin, a G protein-coupled receptor, have previously been limited to apoprotein or opsin forms that do not contain the agonist all-trans-retinal. Two groups now report structures that reveal more details of the transformations involved in rhodopsin activation. Choe et al. solve the X-ray crystal structure of the metarhodopsin II intermediate of the photoreceptor rhodopsin, and Standfuss et al. determine the structure of a constitutively active mutant of rhodopsin bound to a peptide derived from the C-terminus of the G protein transducin. Here the X-ray crystal structure of the metarhodopsin II intermediate of the photoreceptor rhodopsin is solved. Comparison of this structure with previously published structures enabled the proposal of how retinal translocation and rotation induce the conformational changes observed in this structure. G-protein-coupled receptors (GPCRs) are seven transmembrane helix (TM) proteins that transduce signals into living cells by binding extracellular ligands and coupling to intracellular heterotrimeric G proteins (Gαβγ)1. The photoreceptor rhodopsin couples to transducin and bears its ligand 11-cis-retinal covalently bound via a protonated Schiff base to the opsin apoprotein2. Absorption of a photon causes retinal cis/trans isomerization and generates the agonist all-trans-retinal in situ. After early photoproducts, the active G-protein-binding intermediate metarhodopsin II (Meta II) is formed, in which the retinal Schiff base is still intact but deprotonated. Dissociation of the proton from the Schiff base breaks a major constraint in the protein and enables further activating steps, including an outward tilt of TM6 and formation of a large cytoplasmic crevice for uptake of the interacting C terminus of the Gα subunit3,4,5. Owing to Schiff base hydrolysis, Meta II is short-lived and notoriously difficult to crystallize. We therefore soaked opsin crystals with all-trans-retinal to form Meta II, presuming that the crystal’s high concentration of opsin in an active conformation (Ops*)6,7 may facilitate all-trans-retinal uptake and Schiff base formation. Here we present the 3.0 A and 2.85 A crystal structures, respectively, of Meta II alone or in complex with an 11-amino-acid C-terminal fragment derived from Gα (GαCT2). GαCT2 binds in a large crevice at the cytoplasmic side, akin to the binding of a similar Gα-derived peptide to Ops* (ref. 7). In the Meta II structures, the electron density from the retinal ligand seamlessly continues into the Lys 296 side chain, reflecting proper formation of the Schiff base linkage. The retinal is in a relaxed conformation and almost undistorted compared with pure crystalline all-trans-retinal. By comparison with early photoproducts we propose how retinal translocation and rotation induce the gross conformational changes characteristic for Meta II. The structures can now serve as models for the large GPCR family.read more
Citations
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Journal ArticleDOI
Crystal structure of the β2 adrenergic receptor-Gs protein complex.
Søren G. F. Rasmussen,Brian T. DeVree,Yaozhong Zou,Andrew C. Kruse,Ka Young Chung,Tong Sun Kobilka,Foon Sun Thian,Pil Seok Chae,Els Pardon,Els Pardon,Diane M. Calinski,Jesper Mosolff Mathiesen,Syed T. A. Shah,Joseph A. Lyons,Martin Caffrey,Samuel H. Gellman,Jan Steyaert,Jan Steyaert,Georgios Skiniotis,William I. Weis,Roger K. Sunahara,Brian K. Kobilka +21 more
TL;DR: This crystal structure represents the first high-resolution view of transmembrane signalling by a GPCR and the most surprising observation is a major displacement of the α-helical domain of Gαs relative to the Ras-like GTPase domain.
Journal ArticleDOI
Molecular signatures of G-protein-coupled receptors.
AJ Venkatakrishnan,Xavier Deupi,Guillaume Lebon,Christopher G. Tate,Gebhard F. X. Schertler,Gebhard F. X. Schertler,M. Madan Babu +6 more
TL;DR: Through a systematic analysis of high-resolution GPCR structures, a conserved network of non-covalent contacts that defines the G PCR fold is uncovered and characteristic features of ligand binding and conformational changes during receptor activation are revealed.
Journal ArticleDOI
Microbial and animal rhodopsins: structures, functions, and molecular mechanisms.
Oliver P. Ernst,David T. Lodowski,Marcus Elstner,Peter Hegemann,Leonid S. Brown,Hideki Kandori +5 more
TL;DR: Rhodopsins found in Eukaryotes, Bacteria, and Archaea consist of opsin apoproteins and a covalently linked retinal which is employed to absorb photons for energy conversion or the initiation of intra- or intercellular signaling.
Journal ArticleDOI
Activation and allosteric modulation of a muscarinic acetylcholine receptor
Andrew C. Kruse,Aaron M. Ring,Aashish Manglik,Jianxin Hu,Kelly Hu,Katrin Eitel,Harald Hübner,Els Pardon,Celine Valant,Patrick M. Sexton,Arthur Christopoulos,Christian C. Felder,Peter Gmeiner,Jan Steyaert,William I. Weis,K. Christopher Garcia,Jürgen Wess,Brian K. Kobilka +17 more
TL;DR: The structure of an agonist-bound, active state of the human M2 muscarinic acetylcholine receptor stabilized by a G-protein mimetic camelid antibody fragment isolated by conformational selection using yeast surface display reveals larger conformational changes in the extracellular region and orthosteric binding site than observed in the active states of the β2AR and rhodopsin.
Journal ArticleDOI
Conical intersection dynamics of the primary photoisomerization event in vision
Dario Polli,Piero Altoè,Oliver Weingart,Katelyn M. Spillane,Cristian Manzoni,Daniele Brida,Gaia Tomasello,G. Orlandi,Philipp Kukura,Richard A. Mathies,Marco Garavelli,Giulio Cerullo +11 more
TL;DR: Ultrafast optical spectroscopy with sub-20-fs time resolution and spectral coverage from the visible to the near-infrared allows us to follow the dynamics leading to the conical intersection in rhodopsin isomerization and finds excellent agreement between the experimental observations and molecular dynamics calculations that involve a true electronic state crossing.
References
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Journal ArticleDOI
Coot: model-building tools for molecular graphics.
Paul Emsley,Kevin Cowtan +1 more
TL;DR: CCP4mg is a project that aims to provide a general-purpose tool for structural biologists, providing tools for X-ray structure solution, structure comparison and analysis, and publication-quality graphics.
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