Engineering Visual Arrestin-1 with Special Functional Characteristics*

Background: Arrestin-1 with enhanced binding to unphosphorylated active rhodopsin (Rh*) has therapeutic potential. Results: Manipulation of the rhodopsin binding surface of arrestin-1 greatly increases its binding to Rh*. Conclusion: Stable arrestin-1 with high binding to Rh* can be engineered with and without the ability to self-associate. Significance: The affinity of arrestin-1 for Rh* and its propensity to oligomerize can be independently changed by targeted mutagenesis. Arrestin-1 preferentially binds active phosphorylated rhodopsin. Previously, a mutant with enhanced binding to unphosphorylated active rhodopsin (Rh*) was shown to partially compensate for lack of rhodopsin phosphorylation in vivo. Here we showed that reengineering of the receptor binding surface of arrestin-1 further improves the binding to Rh* while preserving protein stability. In mammals, arrestin-1 readily self-associates at physiological concentrations. The biological role of this phenomenon can only be elucidated by replacing wild type arrestin-1 in living animals with a non-oligomerizing mutant retaining all other functions. We demonstrate that constitutively monomeric forms of arrestin-1 are sufficiently stable for in vivo expression. We also tested the idea that individual functions of arrestin-1 can be independently manipulated to generate mutants with the desired combinations of functional characteristics. Here we showed that this approach is feasible; stable forms of arrestin-1 with high Rh* binding can be generated with or without the ability to self-associate. These novel molecular tools open the possibility of testing of the biological role of arrestin-1 self-association and pave the way to elucidation of full potential of compensational approach to gene therapy of gain-of-function receptor mutations.

• Publication year

  2012

• Venue

  Journal of Biological Chemistry

• Publication date

  2012-12-17

• Fields of study

  Biology, Medicine, Engineering

• Identifiers
  DOI 10.1074/jbc.M112.445437PMID 23250748PMCID PMC3561558
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Arrestin interactions with G protein-coupled receptors.

  2014cited by this paper
• Role of Receptor-attached Phosphates in Binding of Visual and Non-visual Arrestins to G Protein-coupled Receptors*

  2012cited by this paper
• Manipulation of Very Few Receptor Discriminator Residues Greatly Enhances Receptor Specificity of Non-visual Arrestins*

  2012cited by this paper
• Silent Scaffolds

  2012cited by this paper
• Conformation of receptor-bound visual arrestin

  2012cited by this paper
• Synthetic biology with surgical precision: targeted reengineering of signaling proteins.

  2012cited by this paper
• Arrestin-1 expression level in rods: balancing functional performance and photoreceptor health.

  2011cited by this paper
• Robust self-association is a common feature of mammalian visual arrestin-1.

  2011cited by this paper
• Identification of Arrestin-3-specific Residues Necessary for JNK3 Kinase Activation*

  2011cited by this paper
• Few Residues within an Extensive Binding Interface Drive Receptor Interaction and Determine the Specificity of Arrestin Proteins*

  2011cited by this paper
• A single mutation in arrestin-2 prevents ERK1/2 activation by reducing c-Raf1 binding.

  2011cited by this paper
• Crystal structure of arrestin-3 reveals the basis of the difference in receptor binding between two non-visual subtypes.

  2011cited by this paper
• The functional cycle of visual arrestins in photoreceptor cells.

  2011cited by this paper
• Identification of key factors that reduce the variability of the single photon response

  2011cited by this paper
• Build life to understand it

  2010cited by this paper
• Phosphotyrosine signaling: evolving a new cellular communication system.

  2010cited by this paper
• Kinetics of Rhodopsin Deactivation and Its Role in Regulating Recovery and Reproducibility of Rod Photoresponse

  2010cited by this paper
• Control of Rhodopsin's Active Lifetime by Arrestin-1 Expression in Mammalian Rods

  2010cited by this paper
• Custom-designed proteins as novel therapeutic tools? The case of arrestins

  2010cited by this paper
• Enhanced arrestin facilitates recovery and protects rods lacking rhodopsin phosphorylation.

  2009cited by this paper
• Treatment of Leber Congenital Amaurosis Due to RPE65 Mutations by Ocular Subretinal Injection of Adeno-Associated Virus Gene Vector: Short-Term Results of a Phase I Trial

  2008cited by this paper
• Human gene therapy for RPE65 isomerase deficiency activates the retinoid cycle of vision but with slow rod kinetics

  2008cited by this paper
• A model for the solution structure of the rod arrestin tetramer.

  2008cited by this paper
• Effect of gene therapy on visual function in Leber's congenital amaurosis.

  2008cited by this paper
• Rich Tapestry of G Protein-Coupled Receptor Signaling and Regulatory Mechanisms

  2008cited by this paper
• Opposing effects of inositol hexakisphosphate on rod arrestin and arrestin2 self-association.

  2008cited by this paper
• Safety and efficacy of gene transfer for Leber's congenital amaurosis.

  2008cited by this paper
• Structure and function of the visual arrestin oligomer

  2007cited by this paper
• Each rhodopsin molecule binds its own arrestin

  2007cited by this paper
• Cone arrestin binding to JNK3 and Mdm2: conformational preference and localization of interaction sites

  2007cited by this paper
• Regulation of Arrestin Binding by Rhodopsin Phosphorylation Level*

  2007cited by this paper
• Arrestins: ubiquitous regulators of cellular signaling pathways

  2006cited by this paper
• Arrestin Translocation Is Induced at a Critical Threshold of Visual Signaling and Is Superstoichiometric to Bleached Rhodopsin

  2006cited by this paper
• Visual and Both Non-visual Arrestins in Their “Inactive” Conformation Bind JNK3 and Mdm2 and Relocalize Them from the Nucleus to the Cytoplasm*

  2006cited by this paper
• The structural basis of arrestin-mediated regulation of G-protein-coupled receptors.

  2006cited by this paper
• The Differential Engagement of Arrestin Surface Charges by the Various Functional Forms of the Receptor*

  2006cited by this paper
• Light-dependent redistribution of arrestin in vertebrate rods is an energy-independent process governed by protein-protein interactions.

  2005cited by this paper
• Crystal structure of cone arrestin at 2.3A: evolution of receptor specificity.

  2005cited by this paper
• Arrestin2 expression selectively increases during neural differentiation

  2004cited by this paper
• Mutant G-protein-coupled receptors as a cause of human diseases.

  2004cited by this paper
• Molecular mechanisms of ligand binding, signaling, and regulation within the superfamily of G-protein-coupled receptors: molecular modeling and mutagenesis approaches to receptor structure and function.

  2004cited by this paper
• The Nature of the Arrestin·Receptor Complex Determines the Ultimate Fate of the Internalized Receptor*

  2003cited by this paper
• Concentration-dependent tetramerization of bovine visual arrestin.

  2003cited by this paper
• Transition of Arrestin into the Active Receptor-binding State Requires an Extended Interdomain Hinge*

  2002cited by this paper
• Conservation of the Phosphate-sensitive Elements in the Arrestin Family of Proteins*

  2002cited by this paper
• Arrestin2 and arrestin3 are differentially expressed in the rat brain during postnatal development.

  2002cited by this paper
• Activation, deactivation, and adaptation in vertebrate photoreceptor cells.

  2001cited by this paper
• Crystal structure of beta-arrestin at 1.9 A: possible mechanism of receptor binding and membrane Translocation.

  2001cited by this paper
• Threonine 180 Is Required for G-protein-coupled Receptor Kinase 3- and β-Arrestin 2-mediated Desensitization of the μ-Opioid Receptor in Xenopus Oocytes*

  2001cited by this paper
• Constitutive arrestin-mediated desensitization of a human vasopressin receptor mutant associated with nephrogenic diabetes insipidus.

  2001cited by this paper
• Arrestin: mutagenesis, expression, purification, and functional characterization.

  2000cited by this paper
• An Additional Phosphate-binding Element in Arrestin Molecule

  2000cited by this paper
• [29] Arrestin: Mutagenesis, expression, purification, and functional characterization

  2000cited by this paper
• The 2.8 A crystal structure of visual arrestin: a model for arrestin's regulation.

  1999cited by this paper
• A Model for Arrestin’s Regulation: The 2.8 Å Crystal Structure of Visual Arrestin

  1999cited by this paper
• How Does Arrestin Respond to the Phosphorylated State of Rhodopsin?*

  1999cited by this paper
• Targeted Construction of Phosphorylation-independent β-Arrestin Mutants with Constitutive Activity in Cells*

  1999cited by this paper
• Visual Arrestin Activity May Be Regulated by Self-association*

  1999cited by this paper
• Origin of reproducibility in the responses of retinal rods to single photons.

  1998cited by this paper
• The Selectivity of Visual Arrestin for Light-activated Phosphorhodopsin Is Controlled by Multiple Nonredundant Mechanisms*

  1998cited by this paper
• Defects in the rhodopsin kinase gene in the Oguchi form of stationary night blindness

  1997cited by this paper
• Mechanism of phosphorylation-recognition by visual arrestin and the transition of arrestin into a high affinity binding state.

  1997cited by this paper
• Use of bacteriophage RNA polymerase in RNA synthesis.

  1996cited by this paper
• A homozygous 1–base pair deletion in the arrestin gene is a frequent cause of Oguchi disease in Japanese

  1995cited by this paper
• Constitutive activation of opsin: interaction of mutants with rhodopsin kinase and arrestin.

  1995cited by this paper
• Mechanisms of rhodopsin inactivation in vivo as revealed by a COOH-terminal truncation mutant

  1995cited by this paper
• Visual Arrestin Binding to Rhodopsin

  1995cited by this paper
• Arrestin Interactions with G Protein-coupled Receptors

  1995cited by this paper
• Ocular findings in a family with autosomal dominant retinitis pigmentosa and a frameshift mutation altering the carboxyl terminal sequence of rhodopsin.

  1993influential reference
• Visual arrestin interaction with rhodopsin. Sequential multisite binding ensures strict selectivity toward light-activated phosphorylated rhodopsin.

  1993cited by this paper
• Cell-free expression of visual arrestin. Truncation mutagenesis identifies multiple domains involved in rhodopsin interaction.

  1992cited by this paper
• Responses of retinal rods to single photons.

  1979cited by this paper
• ReportRGS Expression Rate-Limits Recovery of Rod Photoresponses

  year unknowncited by this paper

• The Role of Individual Residues in the N-Terminus of Arrestin-1 in Rhodopsin Binding

  2025cites this paper
• The Functional Role of Gate Loop Residues in Arrestin Binding to GPCRs

  2025cites this paper
• Arrestins: A Small Family of Multi-Functional Proteins

  2024cites this paper
• Expression of Untagged Arrestins in E. coli and Their Purification

  2023cites this paper
• Modulating signalling lifetime to optimise a prototypical animal opsin for optogenetic applications

  2023cites this paper
• Dynamic Nature of Proteins is Critically Important for Their Function: GPCRs and Signal Transducers

  2023cites this paper
• Functional Role of Arrestin-1 Residues Interacting with Unphosphorylated Rhodopsin Elements

  2023cites this paper
• Solo vs. Chorus: Monomers and Oligomers of Arrestin Proteins

  2022cites this paper
• The Role of Arrestin-1 Middle Loop in Rhodopsin Binding

  2022cites this paper
• Structural evidence for visual arrestin priming via complexation of phosphoinositols.

  2021cites this paper
• Lysine in the lariat loop of arrestins does not serve as phosphate sensor

  2020cites this paper
• An arrestin-1 surface opposite of its interface with photoactivated rhodopsin engages with enolase-1

  2020cites this paper
• Targeting arrestin interactions with its partners for therapeutic purposes.

  2020cites this paper
• The finger loop as an activation sensor in arrestin

  2020cites this paper
• Arrestin mutations: Some cause diseases, others promise cure.

  2019cites this paper
• Arrestin-1 engineering facilitates complex stabilization with native rhodopsin

  2019cites this paper
• Arrestins: structural disorder creates rich functionality

  2018influential citation
• Enhanced Mutant Compensates for Defects in Rhodopsin Phosphorylation in the Presence of Endogenous Arrestin-1

  2018cites this paper
• Molecular Defects of the Disease-Causing Human Arrestin-1 C147F Mutant

  2018cites this paper
• GPCR structure, function, drug discovery and crystallography: report from Academia-Industry International Conference (UK Royal Society)

  2018cites this paper
• Differential manipulation of arrestin-3 binding to basal and agonist-activated G protein-coupled receptors.

  2017cites this paper
• Functional role of the three conserved cysteines in the N domain of visual arrestin-1

  2017cites this paper
• Molecular assembly of rhodopsin with G protein-coupled receptor kinases

  2017cites this paper
• Phosphate Sensor and Construction of Phosphorylation-Independent Arrestins

  2017cites this paper
• The arrestin-rhodopsin complex - Biophysical and Structural Characterisation

  2016cites this paper
• G Protein-coupled Receptor Kinases of the GRK4 Protein Subfamily Phosphorylate Inactive G Protein-coupled Receptors (GPCRs)*

  2015cites this paper
• Arrestin Interactions with Rhodopsin in the Squid Visual System

  2015cites this paper
• GPCR structure, function, drug discovery and crystallography: report from Academia-Industry International Conference (UK Royal Society) Chicheley Hall, 1–2 September 2014

  2015cites this paper
• The rhodopsin-arrestin-1 interaction in bicelles.

  2015cites this paper
• Analyzing the roles of multi-functional proteins in cells: The case of arrestins and GRKs

  2015cites this paper
• Beyond traditional pharmacology: new tools and approaches

  2015cites this paper
• How genetic errors in GPCRs affect their function: Possible therapeutic strategies

  2015cites this paper
• Enhanced phosphorylation-independent arrestins and gene therapy.

  2014cites this paper
• Arrestins - Pharmacology and Therapeutic Potential

  2014cites this paper
• Arrestin-3 binds the MAP kinase JNK3α2 via multiple sites on both domains.

  2014cites this paper
• Self-association of arrestin family members.

  2014cites this paper
• G Protein-Coupled Receptor Genetics

  2014cites this paper
• Functional map of arrestin-1 at single amino acid resolution

  2014cites this paper
• Arrestin Expression in E. coli and Purification

  2014cites this paper
• Analysis and Stabilization of the Arrestin−Rhodopsin Signaling Complex by Scanning Mutagenesis

  2014influential citation
• Design of Super-arrestins for Gene Therapy of Diseases Associated with Excessive Signaling of G Protein-Coupled Receptors

  2014cites this paper
• Therapeutic potential of small molecules and engineered proteins.

  2014cites this paper
• Rapid degeneration of rod photoreceptors expressing self-association-deficient arrestin-1 mutant.

  2013cites this paper
• Constitutively active rhodopsin mutants causing night blindness are effectively phosphorylated by GRKs but differ in arrestin-1 binding.

  2013cites this paper
• Examining the effect of pH on the structure and stability of CLIC1 with E228L and E85L CLIC1 variants

  2013cites this paper
• Critical Role of the Central 139-Loop in Stability and Binding Selectivity of Arrestin-1*

  2013influential citation
• Arrestin-1 Stability and Binding Selectivity of Critical Role of the Central 139-Loop in Signal Transduction :

  2013influential citation