title: Formation and decay of the arrestin·rhodopsin complex in native disc membranes.
abstract: In the G protein-coupled receptor rhodopsin, light-induced cis/trans isomerization of the retinal ligand triggers a series of distinct receptor states culminating in the active Metarhodopsin II (Meta II) state, which binds and activates the G protein transducin (Gt). Long before Meta II decays into the aporeceptor opsin and free all-trans-retinal, its signaling is quenched by receptor phosphorylation and binding of the protein arrestin-1, which blocks further access of Gt to Meta II. Although recent crystal structures of arrestin indicate how it might look in a precomplex with the phosphorylated receptor, the transition into the high affinity complex is not understood. Here we applied Fourier transform infrared spectroscopy to monitor the interaction of arrestin-1 and phosphorylated rhodopsin in native disc membranes. By isolating the unique infrared signature of arrestin binding, we directly observed the structural alterations in both reaction partners. In the high affinity complex, rhodopsin adopts a structure similar to Gt-bound Meta II. In arrestin, a modest loss of β-sheet structure indicates an increase in flexibility but is inconsistent with a large scale structural change. During Meta II decay, the arrestin-rhodopsin stoichiometry shifts from 1:1 to 1:2. Arrestin stabilizes half of the receptor population in a specific Meta II protein conformation, whereas the other half decays to inactive opsin. Altogether these results illustrate the distinct binding modes used by arrestin to interact with different functional forms of the receptor.
authors.affiliation: From the Institut für Medizinische Physik und Biophysik, Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany and.
authors.forename: Florent
authors.initials: F
authors.lastname: Beyrière
authors.affiliation: From the Institut für Medizinische Physik und Biophysik, Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany and.
authors.forename: Martha E
authors.initials: ME
authors.lastname: Sommer
authors.affiliation: From the Institut für Medizinische Physik und Biophysik, Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany and.
authors.forename: Michal
authors.initials: M
authors.lastname: Szczepek
authors.affiliation: From the Institut für Medizinische Physik und Biophysik, Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany and Zentrum für Biophysik und Bioinformatik (BPI) and.
authors.forename: Franz J
authors.initials: FJ
authors.lastname: Bartl
authors.affiliation: From the Institut für Medizinische Physik und Biophysik, Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany and Zentrum für Biophysik und Bioinformatik (BPI) and.
authors.forename: Klaus Peter
authors.initials: KP
authors.lastname: Hofmann
authors.affiliation: From the Institut für Medizinische Physik und Biophysik, Charité-Universitätsmedizin Berlin, 10117 Berlin, Germany and.
authors.forename: Martin
authors.initials: M
authors.lastname: Heck
authors.affiliation: Institut für Biologie, Experimentelle Biophysik, Humboldt-Universität zu Berlin, 10115 Berlin, Germany eglof.ritter@hu-berlin.de.
authors.forename: Eglof
authors.initials: E
authors.lastname: Ritter
mesh_terms: D000818:Animals; D019393:Arrestin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D002462:Cell Membrane / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D055550:Protein Stability; D012243:Rhodopsin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D019393:Arrestin; D046912:Multiprotein Complexes; D012243:Rhodopsin
keywords: G protein-coupled receptor (GPCR); arrestin; infrared spectroscopy (IR spectroscopy); rhodopsin; signal transduction; spectroscopy
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