Structure and Orientation of Ligands Bound to Membrane Proteins Are Reflected by Residual Dipolar Couplings in Solution NMR Measurements

Abstract

Nuclear spins remember the past! The bound conformation of small ligands that are in fast exchange between a free and a membrane‐protein‐bound form can be obtained from solution NMR spectroscopy experiments on the free ligand (see figure). Transferred dipolar couplings characterize both the structure and orientation of a rhodopsin‐bound peptide that mimics a receptor binding region of the G protein transducin. A model for the mutual orientation of rhodopsin and transducin in the active complex is proposed.

Modern High Resolution NMR for the Study of Structure, Dynamics and Interactions of Biological Macromolecules

“…The peptide binds most likely to a site on the third cytoplasmic loop of photo-activated rhodopsin (orange) [282]. (reproduced from [278] with kind permission of Wiley-VCH).…”

Modern High Resolution NMR for the Study of Structure, Dynamics and Interactions of Biological Macromolecules

“…The peptide binds most likely to a site on the third cytoplasmic loop of photo-activated rhodopsin (orange) [282]. (reproduced from [278] with kind permission of Wiley-VCH).…”

Analysis of interactions responsible for vasopressin binding to human neurohypophyseal hormone receptors—molecular dynamics study of the activated receptor–vasopressin–Gα systems

“…These conformational rearrangements eventually result in conversion from rhodopsin to meta rhodopsin (Meta II, MII). It is also known that the C-terminal peptides Gt α (340-350) [41,42] and Gt γ (60-72)farnesyl [43] of transducin α and γ segments, respectively, independently stabilize Meta II [44][45][46]. Gt α (340-350) makes an α-helical extension of the last α5 Gt α helix, potentially fitting into the MII cavity on its cytosolic side [47][48][49].…”

Modern High Resolution NMR for the Study of Structure, Dynamics and Interactions of Biological Macromolecules

“…The peptide binds most likely to a site on the third cytoplasmic loop of photo-activated rhodopsin (orange) [282]. (reproduced from [278] with kind permission of Wiley-VCH). lar information on the target-bound conformation of small molecules in rapidly exchanging complexes of large target and small ligand molecules [274][275][276].…”

“…Transferred dipolar couplings (TrDCs) can be measured on the free ligand in case of rapid exchange of the ligand between a target-bound, macroscopically aligned state and the free form in solution [277]. Measured TrDCs are a residence time-weighted average of the couplings in the two states and contain valuable information on structure and orientation of the target-bound ligand [278]. TrDCs in concert with TrNOEs have been used to determine the high resolution structure of a peptide bound to photo-activated rhodopsin, a G protein-coupled receptor, in its natural membrane environment [279] (see Fig.…”

Modern High Resolution NMR for the Study of Structure, Dynamics and Interactions of Biological Macromolecules

“…The peptide binds most likely to a site on the third cytoplasmic loop of photo-activated rhodopsin (orange) [282]. (reproduced from [278] with kind permission of Wiley-VCH).…”

Analysis of interactions responsible for vasopressin binding to human neurohypophyseal hormone receptors—molecular dynamics study of the activated receptor–vasopressin–Gα systems

“…These conformational rearrangements eventually result in conversion from rhodopsin to meta rhodopsin (Meta II, MII). It is also known that the C-terminal peptides Gt α (340-350) [41,42] and Gt γ (60-72)farnesyl [43] of transducin α and γ segments, respectively, independently stabilize Meta II [44][45][46]. Gt α (340-350) makes an α-helical extension of the last α5 Gt α helix, potentially fitting into the MII cavity on its cytosolic side [47][48][49].…”

Modern High Resolution NMR for the Study of Structure, Dynamics and Interactions of Biological Macromolecules

“…The peptide binds most likely to a site on the third cytoplasmic loop of photo-activated rhodopsin (orange) [282]. (reproduced from [278] with kind permission of Wiley-VCH). lar information on the target-bound conformation of small molecules in rapidly exchanging complexes of large target and small ligand molecules [274][275][276].…”

“…Transferred dipolar couplings (TrDCs) can be measured on the free ligand in case of rapid exchange of the ligand between a target-bound, macroscopically aligned state and the free form in solution [277]. Measured TrDCs are a residence time-weighted average of the couplings in the two states and contain valuable information on structure and orientation of the target-bound ligand [278]. TrDCs in concert with TrNOEs have been used to determine the high resolution structure of a peptide bound to photo-activated rhodopsin, a G protein-coupled receptor, in its natural membrane environment [279] (see Fig.…”

Modern High Resolution NMR for the Study of Structure, Dynamics and Interactions of Biological Macromolecules

“…The peptide binds most likely to a site on the third cytoplasmic loop of photo-activated rhodopsin (orange) [282]. (reproduced from [278] with kind permission of Wiley-VCH).…”

Analysis of interactions responsible for vasopressin binding to human neurohypophyseal hormone receptors—molecular dynamics study of the activated receptor–vasopressin–Gα systems

“…These conformational rearrangements eventually result in conversion from rhodopsin to meta rhodopsin (Meta II, MII). It is also known that the C-terminal peptides Gt α (340-350) [41,42] and Gt γ (60-72)farnesyl [43] of transducin α and γ segments, respectively, independently stabilize Meta II [44][45][46]. Gt α (340-350) makes an α-helical extension of the last α5 Gt α helix, potentially fitting into the MII cavity on its cytosolic side [47][48][49].…”

Modern High Resolution NMR for the Study of Structure, Dynamics and Interactions of Biological Macromolecules

“…The peptide binds most likely to a site on the third cytoplasmic loop of photo-activated rhodopsin (orange) [282]. (reproduced from [278] with kind permission of Wiley-VCH). lar information on the target-bound conformation of small molecules in rapidly exchanging complexes of large target and small ligand molecules [274][275][276].…”

“…Transferred dipolar couplings (TrDCs) can be measured on the free ligand in case of rapid exchange of the ligand between a target-bound, macroscopically aligned state and the free form in solution [277]. Measured TrDCs are a residence time-weighted average of the couplings in the two states and contain valuable information on structure and orientation of the target-bound ligand [278]. TrDCs in concert with TrNOEs have been used to determine the high resolution structure of a peptide bound to photo-activated rhodopsin, a G protein-coupled receptor, in its natural membrane environment [279] (see Fig.…”