Interaction of a G Protein-coupled Receptor with a G Protein-derived Peptide Induces Structural Changes in both Peptide and Receptor: A Fourier-transform Infrared Study Using Isotopically Labeled Peptides

Vogel, Reiner; Martell, Swetlana; Mahalingam, Mohana; Engelhard, Martin; Siebert, Friedrich

doi:10.1016/j.jmb.2006.12.019

Cited by 19 publications

(24 citation statements)

References 34 publications

(55 reference statements)

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“…the positive band at 1659 cm -1 , Figure 2B). To be able to directly compare the modes of interaction between the arrestin finger loop and the deprotonated or protonated receptor species (R*·ArrFL-1 and R*H + ·ArrFL-1), we calculated the so-called peptide binding spectra (PBS), which have been used repeatedly to characterize conformational changes and new molecular interactions formed due to peptide binding (38,39). FTIR difference spectra were measured in the presence and absence of 20 mM ArrFL-1 peptide (∆(w/ ArrFL-1) and ∆(w/o ArrFL-1), gray and black lines in Figure 3 and 4, respectively).…”

Section: Arrfl-1/ph Dual Titration Assay -Inmentioning

confidence: 99%

“…Under these conditions E3.49 is still deprotonated. Binding of Gα C-terminal peptide to R*H + has been shown to stabilize one specific substate out of the receptor ensemble and the concomitant structuring of the peptide leads to a distinct negative band around 1530 cm -1 (38,39), likely caused by hydrophobic interaction between the peptide and the inner faces of TM5 and TM6 (2). We observe a very similar spectral feature in the PBS of R*·ArrFL-1 ( Figure 3, blue line), which suggests that there is a hydrophobic stabilization of ArrFL-1 occurring in the deprotonated binding mode, similar to what has been observed for GαCT.…”

Section: Arrfl-1/ph Dual Titration Assay -Inmentioning

confidence: 99%

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The arrestin-1 finger loop interacts with two distinct conformations of active rhodopsin

Elgeti

Kazmin

Rose

et al. 2018

Journal of Biological Chemistry

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Signaling of the prototypical G protein coupled receptor (GPCR) rhodopsin through its cognate G protein transducin (G t ) is quenched when arrestin binds to the activated receptor. Although the overall architecture of the rhodopsin-arrestin complex is known, many questions regarding its specificity remain unresolved. Here, using FTIR difference spectroscopy and a dual pH/ peptide titration assay, we show that rhodopsin maintains certain flexibility upon binding the "finger loop" of visual arrestin (prepared as synthetic peptide ArrFL-1). We found that two distinct complexes can be stabilized depending on the protonation state of E3.49 in the conserved (D)ERY motif. Both complexes exhibit different interaction modes and affinities of ArrFL-1 binding. The plasticity of the receptor within the rhodopsin/ArrFL-1 complex stands in contrast to the complex with the C-terminus of the G t α-subunit (GαCT), which stabilizes only one specific substate out of the conformational ensemble. However, GαCT binding and both ArrFL-1 binding modes involve a direct interaction to conserved R3.50, as determined by site-directed mutagenesis. Our findings highlight the importance of receptor conformational flexibility and cytoplasmic proton uptake for modulation of rhodopsin signaling and thereby extend the picture provided by crystal structures of the rhodopsin/arrestin and rhodopsin/ArrFL-1 complexes. Furthermore, the two binding modes of ArrFL-1 identified here involve motifs of conserved amino acids, which indicates that our results may have elucidated a common modulation mechanism of class A GPCR-G protein/-arrestin signaling.Unlike other class A GPCRs the photoreceptor rhodopsin contains the light-sensitive cofactor retinal as a ligand, which is covalently bound to the opsin apoprotein. Upon light-induced cis/trans isomerization of retinal and subsequent alterations within the binding pocket, the signal propagates via GPCR-conserved interhelical networks towards the cytoplasmic surface of the receptor, where major structural rearrangements take place. The most prominent structural change is the rotational outward tilt of transmembrane (TM) helix 6, which opens a cytoplasmic binding crevice and thus constitutes the main prerequisite for binding and activation of signaling via the G protein transducin (1, 2). Even though first identified in rhodopsin, for many other GPCRs homologous structural changes have been shown to exist, and a common structural and functional framework for GPCRs is being developed (3).

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Section: Arrfl-1/ph Dual Titration Assay -Inmentioning

confidence: 99%

Section: Arrfl-1/ph Dual Titration Assay -Inmentioning

confidence: 99%

The arrestin-1 finger loop interacts with two distinct conformations of active rhodopsin

Elgeti

Kazmin

Rose

et al. 2018

Journal of Biological Chemistry

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“…More recently, FTIR (Fourier-transform infrared resonance) spectroscopic studies involving binding of the G t C-terminal peptide to Rho* demonstrated clear differences between the active site conformation of Meta II stabilized by this peptide and the active-site conformation stabilized instead by the agonist, all- trans -retinal [25]. These results strongly indicate that allosteric changes in Rho, as well as in G t , are important in the regulation of photoactivation and mediation of the light signal.…”

Section: Complexes Between Rho* Intermediates and Gtmentioning

confidence: 99%

Complexes between photoactivated rhodopsin and transducin: progress and questions

Jastrzębska

Tsybovsky

Palczewski

2010

Biochemical Journal

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Activation of GPCRs (G-protein-coupled receptors) leads to conformational changes that ultimately initiate signal transduction. Activated GPCRs transiently combine with and activate heterotrimeric G-proteins resulting in GTP replacement of GDP on the G-protein α subunit. Both the detailed structural changes essential for productive GDP/GTP exchange on the G-protein α subunit and the structure of the GPCR–G-protein complex itself have yet to be elucidated. Nevertheless, transient GPCR–G-protein complexes can be trapped by nucleotide depletion, yielding an empty-nucleotide G-protein–GPCR complex that can be isolated. Whereas early biochemical studies indicated formation of a complex between G-protein and activated receptor only, more recent results suggest that G-protein can bind to pre-activated states of receptor or even couple transiently to non-activated receptor to facilitate rapid responses to stimuli. Efficient and reproducible formation of physiologically relevant, conformationally homogenous GPCR–G-protein complexes is a prerequisite for structural studies designed to address these possibilities.

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“…2B, gray) that was assigned to conformational changes of rhodopsin induced by peptide binding (41). Formation of the C-terminal reverse turn (C-cap) in the arrestin finger loop (42) might also contribute to this band.…”

Section: Methodsmentioning

confidence: 99%

Formation and Decay of the Arrestin·Rhodopsin Complex in Native Disc Membranes

Beyrière

Sommer

Szczepek

et al. 2015

Journal of Biological Chemistry

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Background:Arrestins regulate the signaling of rhodopsin-like G protein-coupled receptors. Results: We isolated the infrared spectra of rhodopsin⅐arrestin-1 complex formation. Conclusion: Complex formation stabilizes the active receptor and is accompanied by ␤-sheet loss. During decay, arrestin stabilizes only half of the receptor population in the active form. Significance: Our new approach extends knowledge from x-ray structures and other recent spectroscopic studies.

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Interaction of a G Protein-coupled Receptor with a G Protein-derived Peptide Induces Structural Changes in both Peptide and Receptor: A Fourier-transform Infrared Study Using Isotopically Labeled Peptides

Cited by 19 publications

References 34 publications

The arrestin-1 finger loop interacts with two distinct conformations of active rhodopsin

The arrestin-1 finger loop interacts with two distinct conformations of active rhodopsin

Complexes between photoactivated rhodopsin and transducin: progress and questions

Formation and Decay of the Arrestin·Rhodopsin Complex in Native Disc Membranes

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