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Interaction of GCAP1 with retinal guanylyl cyclase and calcium: sensitivity to fatty acylation
Guanylyl cyclase activating proteins (GCAPs) are calcium/magnesium binding proteins within neuronal calcium sensor proteins group (NCS) of the EF-hand proteins superfamily. GCAPs activate retinal guanylyl cyclase (RetGC) in vertebrate photoreceptors in response to light-dependent fall of the intrace...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2012
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3284189/ https://www.ncbi.nlm.nih.gov/pubmed/22371697 http://dx.doi.org/10.3389/fnmol.2012.00019 |
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author | Peshenko, Igor V. Olshevskaya, Elena V. Dizhoor, Alexander M. |
author_facet | Peshenko, Igor V. Olshevskaya, Elena V. Dizhoor, Alexander M. |
author_sort | Peshenko, Igor V. |
collection | PubMed |
description | Guanylyl cyclase activating proteins (GCAPs) are calcium/magnesium binding proteins within neuronal calcium sensor proteins group (NCS) of the EF-hand proteins superfamily. GCAPs activate retinal guanylyl cyclase (RetGC) in vertebrate photoreceptors in response to light-dependent fall of the intracellular free Ca(2+) concentrations. GCAPs consist of four EF-hand domains and contain N-terminal fatty acylated glycine, which in GCAP1 is required for the normal activation of RetGC. We analyzed the effects of a substitution prohibiting N-myristoylation (Gly2 → Ala) on the ability of the recombinant GCAP1 to co-localize with its target enzyme when heterologously expressed in HEK293 cells. We also compared Ca(2+) binding and RetGC-activating properties of the purified non-acylated G2A mutant and C14:0 acylated GCAP1 in vitro. The G2A GCAP1 expressed with a C-terminal GFP tag was able to co-localize with the cyclase, albeit less efficiently than the wild type, but much less effectively stimulated cyclase activity in vitro. Ca(2+) binding isotherm of the G2A GCAP1 was slightly shifted toward higher free Ca(2+) concentrations and so was Ca(2+) sensitivity of RetGC reconstituted with the G2A mutant. At the same time, myristoylation had little effect on the high-affinity Ca(2+)-binding in the EF-hand proximal to the myristoyl residue in three-dimensional GCAP1 structure. These data indicate that the N-terminal fatty acyl group may alter the activity of EF-hands in the distal portion of the GCAP1 molecule via presently unknown intramolecular mechanism. |
format | Online Article Text |
id | pubmed-3284189 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-32841892012-02-27 Interaction of GCAP1 with retinal guanylyl cyclase and calcium: sensitivity to fatty acylation Peshenko, Igor V. Olshevskaya, Elena V. Dizhoor, Alexander M. Front Mol Neurosci Neuroscience Guanylyl cyclase activating proteins (GCAPs) are calcium/magnesium binding proteins within neuronal calcium sensor proteins group (NCS) of the EF-hand proteins superfamily. GCAPs activate retinal guanylyl cyclase (RetGC) in vertebrate photoreceptors in response to light-dependent fall of the intracellular free Ca(2+) concentrations. GCAPs consist of four EF-hand domains and contain N-terminal fatty acylated glycine, which in GCAP1 is required for the normal activation of RetGC. We analyzed the effects of a substitution prohibiting N-myristoylation (Gly2 → Ala) on the ability of the recombinant GCAP1 to co-localize with its target enzyme when heterologously expressed in HEK293 cells. We also compared Ca(2+) binding and RetGC-activating properties of the purified non-acylated G2A mutant and C14:0 acylated GCAP1 in vitro. The G2A GCAP1 expressed with a C-terminal GFP tag was able to co-localize with the cyclase, albeit less efficiently than the wild type, but much less effectively stimulated cyclase activity in vitro. Ca(2+) binding isotherm of the G2A GCAP1 was slightly shifted toward higher free Ca(2+) concentrations and so was Ca(2+) sensitivity of RetGC reconstituted with the G2A mutant. At the same time, myristoylation had little effect on the high-affinity Ca(2+)-binding in the EF-hand proximal to the myristoyl residue in three-dimensional GCAP1 structure. These data indicate that the N-terminal fatty acyl group may alter the activity of EF-hands in the distal portion of the GCAP1 molecule via presently unknown intramolecular mechanism. Frontiers Media S.A. 2012-02-22 /pmc/articles/PMC3284189/ /pubmed/22371697 http://dx.doi.org/10.3389/fnmol.2012.00019 Text en Copyright © 2012 Peshenko, Olshevskaya and Dizhoor. http://www.frontiersin.org/licenseagreement This is an open-access article distributed under the terms of the Creative Commons Attribution Non Commercial License, which permits non-commercial use, distribution, and reproduction in other forums, provided the original authors and source are credited. |
spellingShingle | Neuroscience Peshenko, Igor V. Olshevskaya, Elena V. Dizhoor, Alexander M. Interaction of GCAP1 with retinal guanylyl cyclase and calcium: sensitivity to fatty acylation |
title | Interaction of GCAP1 with retinal guanylyl cyclase and calcium: sensitivity to fatty acylation |
title_full | Interaction of GCAP1 with retinal guanylyl cyclase and calcium: sensitivity to fatty acylation |
title_fullStr | Interaction of GCAP1 with retinal guanylyl cyclase and calcium: sensitivity to fatty acylation |
title_full_unstemmed | Interaction of GCAP1 with retinal guanylyl cyclase and calcium: sensitivity to fatty acylation |
title_short | Interaction of GCAP1 with retinal guanylyl cyclase and calcium: sensitivity to fatty acylation |
title_sort | interaction of gcap1 with retinal guanylyl cyclase and calcium: sensitivity to fatty acylation |
topic | Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3284189/ https://www.ncbi.nlm.nih.gov/pubmed/22371697 http://dx.doi.org/10.3389/fnmol.2012.00019 |
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