Rhodopsin Interactions
Published by Anonymous on 2007/9/30 (2607 reads)
1: Methods Mol Biol. 2004;261:93-112.
Measuring rhodopsin-G-protein interactions by surface plasmon resonance.
Northup J.
Laboratory of Cellular Biology, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Rockville, MD, USA.
G-protein-coupled receptors (GPCRs) initiate a variety of cellular responses to diverse array of extracellular stimuli. Surface plasmon resonance detection offers a powerful approach to the study of protein-protein interactions in real time. In this chapter we outline procedures for the immobilization of the prototype GPCR structure, rhodopsin or the G-protein alpha and betagamma subunits, for analysis of the molecular interactions initiating G-protein signaling. The attachment of rhodopsin via its extracellular carbohydrate residues provides a convenient, and universally applicable, procedure for GPCR immobilization in a form that retains full biochemical activity and ability to interact with intracellular signaling components. SPR detection then allows for the analysis of the kinetic and equilibrium binding properties of the immobilized receptor with G-protein subunits and potentially other interacting molecules.
Publication Types:
Research Support, U.S. Gov't, P.H.S.
Review
PMID: 15064451 [PubMed - indexed for MEDLINE]
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2: Cell Mol Neurobiol. 1991 Dec;11(6):563-78.
Molecular interactions between the photoreceptor G protein and rhodopsin.
Hamm HE.
Department of Physiology and Biophysics, University of Illinois College of Medicine, Chicago 60680.
1. The visual transduction system of the vertebrate retina is a well-studied model for biochemical and molecular studies of signal transduction. The structure and function of rhodopsin, a prototypical G protein-coupled receptor, and transducin or Gt, the photoreceptor G protein, have been particularly well studied. Mechanisms of rhodopsin-Gt interaction are discussed in this review. 2. The visual pigment rhodopsin contains a chromophore, and thus conformational changes leading to activation can be monitored spectroscopically. A model of the conformational changes in the activated receptor is presented based on biophysical and biochemical data. 3. The current information on sites of interaction on receptors and cognate G proteins is summarized. Studies using synthetic peptides from amino acid sequences corresponding to Gt and rhodopsin have provided information on the sites of rhodopsin-Gt interaction. Synthetic peptides from the carboxyl terminal region of alpha t mimic Gt by stabilizing the active conformation of rhodopsin, Metarhodopsin II. 4. The conformation of one such peptide when it is bound to Metarhodopsin II was determined by 2D NMR. The model based on the NMR data was tested using peptide analogs predicted to stabilize or break the structure. These studies yield molecular insight into why toxin-treated and mutant G proteins are uncoupled from receptors.
Publication Types:
Review
PMID: 1782650 [PubMed - indexed for MEDLINE]
Measuring rhodopsin-G-protein interactions by surface plasmon resonance.
Northup J.
Laboratory of Cellular Biology, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Rockville, MD, USA.
G-protein-coupled receptors (GPCRs) initiate a variety of cellular responses to diverse array of extracellular stimuli. Surface plasmon resonance detection offers a powerful approach to the study of protein-protein interactions in real time. In this chapter we outline procedures for the immobilization of the prototype GPCR structure, rhodopsin or the G-protein alpha and betagamma subunits, for analysis of the molecular interactions initiating G-protein signaling. The attachment of rhodopsin via its extracellular carbohydrate residues provides a convenient, and universally applicable, procedure for GPCR immobilization in a form that retains full biochemical activity and ability to interact with intracellular signaling components. SPR detection then allows for the analysis of the kinetic and equilibrium binding properties of the immobilized receptor with G-protein subunits and potentially other interacting molecules.
Publication Types:
Research Support, U.S. Gov't, P.H.S.
Review
PMID: 15064451 [PubMed - indexed for MEDLINE]
--------------------------------------------------------------------------------
2: Cell Mol Neurobiol. 1991 Dec;11(6):563-78.
Molecular interactions between the photoreceptor G protein and rhodopsin.
Hamm HE.
Department of Physiology and Biophysics, University of Illinois College of Medicine, Chicago 60680.
1. The visual transduction system of the vertebrate retina is a well-studied model for biochemical and molecular studies of signal transduction. The structure and function of rhodopsin, a prototypical G protein-coupled receptor, and transducin or Gt, the photoreceptor G protein, have been particularly well studied. Mechanisms of rhodopsin-Gt interaction are discussed in this review. 2. The visual pigment rhodopsin contains a chromophore, and thus conformational changes leading to activation can be monitored spectroscopically. A model of the conformational changes in the activated receptor is presented based on biophysical and biochemical data. 3. The current information on sites of interaction on receptors and cognate G proteins is summarized. Studies using synthetic peptides from amino acid sequences corresponding to Gt and rhodopsin have provided information on the sites of rhodopsin-Gt interaction. Synthetic peptides from the carboxyl terminal region of alpha t mimic Gt by stabilizing the active conformation of rhodopsin, Metarhodopsin II. 4. The conformation of one such peptide when it is bound to Metarhodopsin II was determined by 2D NMR. The model based on the NMR data was tested using peptide analogs predicted to stabilize or break the structure. These studies yield molecular insight into why toxin-treated and mutant G proteins are uncoupled from receptors.
Publication Types:
Review
PMID: 1782650 [PubMed - indexed for MEDLINE]
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