Abstract
G-protein-coupled receptors (GPCRs) are essential mediators of information transfer in eukaryotic cells. Interactions between GPCRs and their binding partners modulate the signaling process. For example, the interaction between GPCR and cognate G protein initiates the signal, while the interaction with cognate arrestin terminates G-protein-mediated signaling. In visual signal transduction, arrestin-1 selectively binds to the phosphorylated light-activated GPCR rhodopsin to terminate rhodopsin signaling. Under physiological conditions, the rhodopsin-arrestin-1 interaction occurs in highly specialized disk membrane in which rhodopsin resides. This membrane is replaced with mimetics when working with purified proteins. While detergents are commonly used as membrane mimetics, most detergents denature arrestin-1, preventing biochemical studies of this interaction. In contrast, bicelles provide a suitable alternative medium. An advantage of bicelles is that they contain lipids, which have been shown to be necessary for normal rhodopsin-arrestin-1 interaction. Here we describe how to reconstitute rhodopsin into bicelles, and how bicelle properties affect the rhodopsin-arrestin-1 interaction.
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References
Wilden U, Hall SW, Kühn H (1986) Phosphodiesterase activation by photoexcited rhodopsin is quenched when rhodopsin is phosphorylated and binds the intrinsic 48-kDa protein of rod outer segments. Proc Natl Acad Sci U S A 83:1174–1178
Krupnick JG, Gurevich VV, Benovic JL (1997) Mechanism of quenching of phototransduction. Binding competition between arrestin and transducin for phosphorhodopsin. J Biol Chem 272:18125–18131
Jastrzebska B, Debinski A, Filipek S et al (2011) Role of membrane integrity on G protein-coupled receptors: rhodopsin stability and function. Prog Lipid Res 50:267–277
Brown MF (1994) Modulation of rhodopsin function by properties of the membrane bilayer. Chem Phys Lipids 73:159–180
Vishnivetskiy SA, Raman D, Wei J et al (2007) Regulation of arrestin binding by rhodopsin phosphorylation level. J Biol Chem 282:32075–32083
Bayburt TH, Vishnivetskiy SA, McLean MA et al (2011) Monomeric rhodopsin is sufficient for normal rhodopsin kinase (GRK1) phosphorylation and arrestin-1 binding. J Biol Chem 286:1420–1428
Gurevich VV, Benovic JL (1993) Visual arrestin interaction with rhodopsin. Sequential multisite binding ensures strict selectivity toward light-activated phosphorylated rhodopsin. J Biol Chem 268:11628–11638
Gurevich VV (1998) The selectivity of visual arrestin for light-activated phosphorhodopsin is controlled by multiple nonredundant mechanisms. J Biol Chem 273:15501–15506
Gurevich VV, Benovic JL (1995) Visual arrestin binding to rhodopsin: diverse functional roles of positively charged residues within the phosphorylation-recognition region of arrestin. J Biol Chem 270:6010–6016
Gurevich VV, Benovic JL (1997) Mechanism of phosphorylation-recognition by visual arrestin and the transition of arrestin into a high affinity binding state. Mol Pharmacol 51:161–169
Gurevich VV, Gurevich EV (2004) The molecular acrobatics of arrestin activation. Trends Pharmacol Sci 25:105–111
Hirsch JA, Schubert C, Gurevich VV et al (1999) The 2.8 angstrom crystal structure of visual arrestin: a model for arrestin’s regulation. Cell 97:257–269
Granzin J, Cousin A, Weirauch M et al (2012) Crystal structure of p44, a constitutively active splice variant of visual arrestin. J Mol Biol 416:611–618
Kim YJ, Hofmann KP, Ernst OP et al (2013) Crystal structure of pre-activated arrestin p44. Nature 497:142–146
Zhuang TD, Chen QY, Cho MK et al (2013) Involvement of distinct arrestin-1 elements in binding to different functional forms of rhodopsin. Proc Natl Acad Sci U S A 110:942–947
Vishnivetskiy SA, Schubert C, Climaco GC et al (2000) An additional phosphate-binding element in arrestin molecule: implications for the mechanism of arrestin activation. J Biol Chem 275:41049–41057
Vishnivetskiy SA, Paz CL, Schubert C et al (1999) How does arrestin respond to the phosphorylated state of rhodopsin? J Biol Chem 274:11451–11454
Vishnivetskiy SA, Francis DJ, Van Eps N et al (2010) The role of arrestin alpha-helix I in receptor binding. J Mol Biol 395:42–54
Hanson SM, Francis DJ, Vishnivetskiy SA et al (2006) Differential interaction of spin-labeled arrestin with inactive and active phosphorhodopsin. Proc Natl Acad Sci U S A 103:4900–4905
Kim M, Vishnivetskiy SA, Van Eps N et al (2012) Conformation of receptor-bound visual arrestin. Proc Natl Acad Sci U S A 109:18407–18412
Ostermaier MK, Peterhans C, Jaussi R et al (2014) Functional map of arrestin-1 at single amino acid resolution. Proc Natl Acad Sci U S A 111:1825–1830
Vishnivetskiy SA, Baameur F, Findley KR et al (2013) Critical role of the central 139-loop in stability and binding selectivity of arrestin-1. J Biol Chem 288:11741–11750
Vishnivetskiy SA, Chen Q, Palazzo MC et al (2013) Engineering visual arrestin-1 with special functional characteristics. J Biol Chem 288:11741–11750
Sanders CR, Hare BJ, Howard KP et al (1994) Magnetically-oriented phospholipid micelles as a tool for the study of membrane-associated molecules. Prog Nucl Magn Reson Spectrosc 26:421–444
Sanders CR, Prosser RS (1998) Bicelles: a model membrane system for all seasons? Struct Folding Des 6:1227–1234
Ujwal R, Bowie JU (2011) Crystallizing membrane proteins using lipidic bicelles. Methods 55:337–341
Durr UHN, Gildenberg M, Ramamoorthy A (2012) The magic of bicelles lights up membrane protein structure. Chem Rev 112:6054–6074
Rasmussen SG, Choi HJ, Rosenbaum DM et al (2007) Crystal structure of the human beta2 adrenergic G-protein-coupled receptor. Nature 450:383–387
Ye WH, Lind J, Eriksson J et al (2014) Characterization of the morphology of fast-tumbling bicelles with varying composition. Langmuir 30:5488–5496
Beaugrand M, Arnold AA, Henin J et al (2014) Lipid concentration and molar ratio boundaries for the use of isotropic bicelles. Langmuir 30:6162–6170
Thompson AA, Liu JJ, Chun E et al (2011) GPCR stabilization using the bicelle-like architecture of mixed sterol-detergent micelles. Methods 55:310–317
Zocher M, Zhang C, Rasmussen SG et al (2012) Cholesterol increases kinetic, energetic, and mechanical stability of the human beta2-adrenergic receptor. Proc Natl Acad Sci U S A 109:E3463–E3472
Rim J, Oprian DD (1995) Constitutive activation of opsin – interaction of mutants with rhodopsin kinase and arrestin. Biochemistry 34:11938–11945
Degrip WJ (1982) Thermal-stability of rhodopsin and opsin in some novel detergents. Methods Enzymol 81:256–265
Reeves PJ, Hwa J, Khorana HG (1999) Structure and function in rhodopsin: kinetic studies of retinal binding to purified opsin mutants in defined phospholipid-detergent mixtures serve as probes of the retinal binding pocket. Proc Natl Acad Sci U S A 96:1927–1931
McKibbin C, Farmer NA, Jeans C et al (2007) Opsin stability and folding: modulation by phospholipid bicelles. J Mol Biol 374:1319–1332
Gurevich VV, Gurevich EV (2008) GPCR monomers and oligomers: it takes all kinds. Trends Neurosci 31:74–81
Gurevich VV, Gurevich EV (2008) How and why do GPCRs dimerize? Trends Pharmacol Sci 29:234–240
Bayburt TH, Leitz AJ, Xie G et al (2007) Transducin activation by nanoscale lipid bilayers containing one and two rhodopsins. J Biol Chem 282:14875–14881
Whorton MR, Jastrzebska B, Park PSH et al (2008) Efficient coupling of transducin to monomeric rhodopsin in a phospholipid bilayer. J Biol Chem 283:4387–4394
Vishnivetskiy SA, Ostermaierm MK, Singhal A et al (2013) Constitutively active rhodopsin mutants causing night blindness are effectively phosphorylated by GRKs but differ in arrestin-1 binding. Cell Signal 25:2155–2162
Singhal A, Ostermaier MK, Vishnivetskiy SA et al (2013) Insights into congenital night blindness based on the structure of G90D rhodopsin. EMBO Rep 14:520–526
Sommer ME, Smith WC, Farrens DL (2006) Dynamics of arrestin-rhodopsin interactions: acidic phospholipids enable binding of arrestin to purified rhodopsin in detergent. J Biol Chem 281:9407–9417
Gurevich VV, Benovic JL (2000) Arrestin: mutagenesis, expression, purification, and functional characterization. Methods Enzymol 315:422–437
Gurevich VV, Benovic JL (1992) Cell-free expression of visual arrestin. Truncation mutagenesis identifies multiple domains involved in rhodopsin interaction. J Biol Chem 267:21919–21923
Gurevich VV (1996) Use of bacteriophage RNA polymerase in RNA synthesis. In: Kuo LC, Olsen DB, Carroll SS (eds) Methods in enzymology, 275: 382–397
Weigelt J (1998) Single scan, sensitivity- and gradient-enhanced TROSY for multidimensional NMR experiments. J Am Chem 120:10778–10779
Tugarinov V, Kay LE (2005) Methyl groups as probes of structure and dynamics in NMR studies of high-molecular-weight proteins. Chembiochem 6:1567–1577
Delaglio F, Grzesiek S, Vuister GW et al (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6:277–293
Johnson BA (2004) Using NMRView to visualize and analyze the NMR spectra of macromolecules. Methods Mol Biol 278:313–352
Goddard TD, Kneller DG (2008) SPARKY 3. University of California, San Francisco
Palczewski K, Kumasaka T, Hori T et al (2000) Crystal structure of rhodopsin: A G protein-coupled receptor. Science 289:739-745
Choe HW, Kim YJ, Park JH et al (2011) Crystal structure of metarhodopsin II. Nature 471:651–655
Alexander NS, Preininger AM, Kaya AI et al (2014) Energetic analysis of the rhodopsin-G-protein complex links the α5 helix to GDP release. Nat Struct Mol Biol 21:56–63
Singh P, Wang B, Maeda T et al (2008) Structures of rhodopsin kinase in different ligand states reveal key elements involved in G protein-coupled receptor kinase activation. J Biol Chem 283:14053–14062
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Chen, Q. et al. (2015). The Rhodopsin-Arrestin-1 Interaction in Bicelles. In: Jastrzebska, B. (eds) Rhodopsin. Methods in Molecular Biology, vol 1271. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2330-4_6
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DOI: https://doi.org/10.1007/978-1-4939-2330-4_6
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