Light Activation of Rhodopsin: Insights from Molecular Dynamics Simulations Guided by Solid-State NMR Distance Restraints
ABSTRACT Structural restraints provided by solid-state NMR measurements of the metarhodopsin II intermediate are combined with molecular dynamics simulations to help visualize structural changes in the light activation of rhodopsin. Since the timescale for the formation of the metarhodopsin II intermediate (> 1 ms) is beyond that readily accessible by molecular dynamics, we use NMR distance restraints derived from 13C dipolar recoupling measurements to guide the simulations. The simulations yield a working model for how photoisomerization of the 11-cis retinylidene chromophore bound within the interior of rhodopsin is coupled to transmembrane helix motion and receptor activation. The mechanism of activation that emerges is that multiple switches on the extracellular (or intradiscal) side of rhodopsin trigger structural changes that converge to disrupt the ionic lock between helices H3 and H6 on the intracellular side of the receptor.
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ABSTRACT: We investigate the nuclear isotropic shielding constants σ((13)C) and σ((17)O) of isomers of retinoic acid and retinal in gas-phase and in chloroform, acetonitrile, methanol, and water solutions via Monte Carlo simulation and quantum mechanics calculations using the GIAO-B3LYP∕6-311++G(2d,2p) approach. Electronic solute polarization effects due to protic and aprotic solvents are included iteratively and play an important role in the quantitative determination of oxygen shielding constants. Our MP2∕6-31G+(d) results show substantial increases of the dipole moment of both retinal derivatives in solution as compared with the gas-phase results (between 22% and 26% in chloroform and between 55% and 99% in water). For the oxygen atoms the influence of the solute polarization is mild for σ((17)O) of hydroxyl group, even in protic solvents, but it is particularly important for σ((17)O) of carbonyl group. For the latter, there is a sizable increase in the magnitude with increasing solvent polarity. For the carbon atoms, the solvent effects on the σ((13)C) values are in general small, being more appreciable in carbon atoms of the polyene chain than in the carbon atoms of the β-ionone ring and methyl groups. The results also show that isomeric changes on the backbones of the polyene chains have marked influence on the (13)C chemical shifts of carbon atoms near to the structural distortions, in good agreement with the experimental results measured in solution.The Journal of Chemical Physics 09/2013; 139(9):094502. · 3.12 Impact Factor
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ABSTRACT: The past decade has witnessed an impressive expansion of our knowledge of retinal photoreceptor signal transduction and the regulation of the visual cycle required for normal eyesight. Progress in human genetics and next generation sequencing technologies have revealed the complexity behind many inherited retinal diseases. Structural studies have markedly increased our understanding of the visual process. Moreover, technical innovations and improved methodologies in proteomics, macromolecular crystallization and high resolution imaging at different levels set the scene for even greater advances. Pharmacology combined with structural biology of membrane proteins holds great promise for developing innovative accessible therapies for millions robbed of their sight or progressing toward blindness.Current opinion in cell biology 04/2014; 27C:32-43. · 14.15 Impact Factor
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ABSTRACT: Rhodopsin, the mammalian dim-light receptor, is one of the best-characterized G-protein-coupled receptors-a pharmaceutically important class of membrane proteins that has garnered much attention due to the recent availability of structural information. Yet, the mechanism of rhodopsin activation is not fully understood. Here, we use microsecond-scale all-atom molecular dynamics simulations, validated by solid-state 2H NMR spectroscopy, to understand the transition between the dark and metarhodopsin I (Meta I) states. Our analysis of these simulations reveals striking differences in ligand flexibility between the two states; retinal is much more dynamic in Meta I, adopting an elongated conformation similar to that seen in the recent active-like crystal structures. Surprisingly, this elongation corresponds to both a dramatic influx of bulk water into the hydrophobic core of the protein and to a concerted transition in the highly conserved Trp2656.48 residue. In addition, enhanced ligand flexibility upon light activation provides an explanation for the different retinal orientations observed in X-ray crystal structures of active rhodopsin.Biochemistry 12/2013; · 3.38 Impact Factor