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An arrestin-1 surface opposite of its interface with photoactivated rhodopsin engages with enolase-1
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 investiga...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7212649/ https://www.ncbi.nlm.nih.gov/pubmed/32238431 http://dx.doi.org/10.1074/jbc.RA120.013043 |
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author | Miranda, Connie Jaqueline Fernandez, Nicole Kamel, Nader Turner, Daniel Benzenhafer, Del Bolch, Susan N. Andring, Jacob T. McKenna, Robert Smith, W. Clay |
author_facet | Miranda, Connie Jaqueline Fernandez, Nicole Kamel, Nader Turner, Daniel Benzenhafer, Del Bolch, Susan N. Andring, Jacob T. McKenna, Robert Smith, W. Clay |
author_sort | Miranda, Connie Jaqueline |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-7212649 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-72126492020-05-18 An arrestin-1 surface opposite of its interface with photoactivated rhodopsin engages with enolase-1 Miranda, Connie Jaqueline Fernandez, Nicole Kamel, Nader Turner, Daniel Benzenhafer, Del Bolch, Susan N. Andring, Jacob T. McKenna, Robert Smith, W. Clay J Biol Chem Cell Biology 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. American Society for Biochemistry and Molecular Biology 2020-05-08 2020-04-01 /pmc/articles/PMC7212649/ /pubmed/32238431 http://dx.doi.org/10.1074/jbc.RA120.013043 Text en © 2020 Miranda et al. Author's Choice—Final version open access under the terms of the Creative Commons CC-BY license (http://creativecommons.org/licenses/by/4.0) . |
spellingShingle | Cell Biology Miranda, Connie Jaqueline Fernandez, Nicole Kamel, Nader Turner, Daniel Benzenhafer, Del Bolch, Susan N. Andring, Jacob T. McKenna, Robert Smith, W. Clay An arrestin-1 surface opposite of its interface with photoactivated rhodopsin engages with enolase-1 |
title | An arrestin-1 surface opposite of its interface with photoactivated rhodopsin engages with enolase-1 |
title_full | An arrestin-1 surface opposite of its interface with photoactivated rhodopsin engages with enolase-1 |
title_fullStr | An arrestin-1 surface opposite of its interface with photoactivated rhodopsin engages with enolase-1 |
title_full_unstemmed | An arrestin-1 surface opposite of its interface with photoactivated rhodopsin engages with enolase-1 |
title_short | An arrestin-1 surface opposite of its interface with photoactivated rhodopsin engages with enolase-1 |
title_sort | arrestin-1 surface opposite of its interface with photoactivated rhodopsin engages with enolase-1 |
topic | Cell Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7212649/ https://www.ncbi.nlm.nih.gov/pubmed/32238431 http://dx.doi.org/10.1074/jbc.RA120.013043 |
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