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An arrestin-1 surface opposite of its interface with photoactivated rhodopsin engages with enolase-1.

Authors
  • Miranda, Connie Jaqueline1
  • Fernandez, Nicole1
  • Kamel, Nader1
  • Turner, Daniel1
  • Benzenhafer, Del1
  • Bolch, Susan N1
  • Andring, Jacob T2
  • McKenna, Robert2
  • Smith, W Clay3
  • 1 Department of Ophthalmology, University of Florida, Gainesville, Florida 32610.
  • 2 Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, Florida 32610.
  • 3 Department of Ophthalmology, University of Florida, Gainesville, Florida 32610 [email protected]
Type
Published Article
Journal
Journal of Biological Chemistry
Publisher
American Society for Biochemistry and Molecular Biology
Publication Date
May 08, 2020
Volume
295
Issue
19
Pages
6498–6508
Identifiers
DOI: 10.1074/jbc.RA120.013043
PMID: 32238431
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Arrestin-1 is the arrestin family member responsible for inactivation of the G protein-coupled receptor rhodopsin in photoreceptors. Arrestin-1 is also well-known to interact with additional protein partners and to affect other signaling cascades beyond phototransduction. In this study, we investigated one of these alternative arrestin-1 binding partners, the glycolysis enzyme enolase-1, to map the molecular contact sites between these two proteins and investigate how the binding of arrestin-1 affects the catalytic activity of enolase-1. Using fluorescence quench protection of strategically placed fluorophores on the arrestin-1 surface, we observed that arrestin-1 primarily engages enolase-1 along a surface that is opposite of the side of arrestin-1 that binds photoactivated rhodopsin. Using this information, we developed a molecular model of the arrestin-1-enolase-1 complex, which was validated by targeted substitutions of charge-pair interactions. Finally, we identified the likely source of arrestin's modulation of enolase-1 catalysis, showing that selective substitution of two amino acids in arrestin-1 can completely remove its effect on enolase-1 activity while still remaining bound to enolase-1. These findings open up opportunities for examining the functional effects of arrestin-1 on enolase-1 activity in photoreceptors and their surrounding cells. © 2020 Miranda et al.

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