Disulfide Dimerization of Neuronal Calcium Sensor-1: Implications for Zinc and Redox Signaling

Neuronal calcium sensor-1 (NCS-1) is a four-EF-hand ubiquitous signaling protein modulating neuronal function and survival, which participates in neurodegeneration and carcinogenesis. NCS-1 recognizes specific sites on cellular membranes and regulates numerous targets, including G-protein coupled re...

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Autores principales:	Viktoriia E. Baksheeva, Alexey V. Baldin, Arthur O. Zalevsky, Aliya A. Nazipova, Alexey S. Kazakov, Vasiliy I. Vladimirov, Neonila V. Gorokhovets, François Devred, Pavel P. Philippov, Alexandr V. Bazhin, Andrey V. Golovin, Andrey A. Zamyatnin, Dmitry V. Zinchenko, Philipp O. Tsvetkov, Sergei E. Permyakov, Evgeni Yu. Zernii
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	EF-hand NCS family neuronal calcium sensor-1 disulfide dimerization GRK1 zinc Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/49fd7d64d9f04e46b0edbc351e7d15fc
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spelling	oai:doaj.org-article:49fd7d64d9f04e46b0edbc351e7d15fc2021-11-25T17:58:16ZDisulfide Dimerization of Neuronal Calcium Sensor-1: Implications for Zinc and Redox Signaling10.3390/ijms2222126021422-00671661-6596https://doaj.org/article/49fd7d64d9f04e46b0edbc351e7d15fc2021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/22/12602https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Neuronal calcium sensor-1 (NCS-1) is a four-EF-hand ubiquitous signaling protein modulating neuronal function and survival, which participates in neurodegeneration and carcinogenesis. NCS-1 recognizes specific sites on cellular membranes and regulates numerous targets, including G-protein coupled receptors and their kinases (GRKs). Here, with the use of cellular models and various biophysical and computational techniques, we demonstrate that NCS-1 is a redox-sensitive protein, which responds to oxidizing conditions by the formation of disulfide dimer (dNCS-1), involving its single, highly conservative cysteine C38. The dimer content is unaffected by the elevation of intracellular calcium levels but increases to 10–30% at high free zinc concentrations (characteristic of oxidative stress), which is accompanied by accumulation of the protein in punctual clusters in the perinuclear area. The formation of dNCS-1 represents a specific Zn<sup>2+</sup>-promoted process, requiring proper folding of the protein and occurring at redox potential values approaching apoptotic levels. The dimer binds Ca<sup>2+</sup> only in one EF-hand per monomer, thereby representing a unique state, with decreased α-helicity and thermal stability, increased surface hydrophobicity, and markedly improved inhibitory activity against GRK1 due to 20-fold higher affinity towards the enzyme. Furthermore, dNCS-1 can coordinate zinc and, according to molecular modeling, has an asymmetrical structure and increased conformational flexibility of the subunits, which may underlie their enhanced target-binding properties. In HEK293 cells, dNCS-1 can be reduced by the thioredoxin system, otherwise accumulating as protein aggregates, which are degraded by the proteasome. Interestingly, NCS-1 silencing diminishes the susceptibility of Y79 cancer cells to oxidative stress-induced apoptosis, suggesting that NCS-1 may mediate redox-regulated pathways governing cell death/survival in response to oxidative conditions.Viktoriia E. BaksheevaAlexey V. BaldinArthur O. ZalevskyAliya A. NazipovaAlexey S. KazakovVasiliy I. VladimirovNeonila V. GorokhovetsFrançois DevredPavel P. PhilippovAlexandr V. BazhinAndrey V. GolovinAndrey A. ZamyatninDmitry V. ZinchenkoPhilipp O. TsvetkovSergei E. PermyakovEvgeni Yu. ZerniiMDPI AGarticleEF-handNCS familyneuronal calcium sensor-1disulfide dimerizationGRK1zincBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 12602, p 12602 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	EF-hand NCS family neuronal calcium sensor-1 disulfide dimerization GRK1 zinc Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	EF-hand NCS family neuronal calcium sensor-1 disulfide dimerization GRK1 zinc Biology (General) QH301-705.5 Chemistry QD1-999 Viktoriia E. Baksheeva Alexey V. Baldin Arthur O. Zalevsky Aliya A. Nazipova Alexey S. Kazakov Vasiliy I. Vladimirov Neonila V. Gorokhovets François Devred Pavel P. Philippov Alexandr V. Bazhin Andrey V. Golovin Andrey A. Zamyatnin Dmitry V. Zinchenko Philipp O. Tsvetkov Sergei E. Permyakov Evgeni Yu. Zernii Disulfide Dimerization of Neuronal Calcium Sensor-1: Implications for Zinc and Redox Signaling
description	Neuronal calcium sensor-1 (NCS-1) is a four-EF-hand ubiquitous signaling protein modulating neuronal function and survival, which participates in neurodegeneration and carcinogenesis. NCS-1 recognizes specific sites on cellular membranes and regulates numerous targets, including G-protein coupled receptors and their kinases (GRKs). Here, with the use of cellular models and various biophysical and computational techniques, we demonstrate that NCS-1 is a redox-sensitive protein, which responds to oxidizing conditions by the formation of disulfide dimer (dNCS-1), involving its single, highly conservative cysteine C38. The dimer content is unaffected by the elevation of intracellular calcium levels but increases to 10–30% at high free zinc concentrations (characteristic of oxidative stress), which is accompanied by accumulation of the protein in punctual clusters in the perinuclear area. The formation of dNCS-1 represents a specific Zn<sup>2+</sup>-promoted process, requiring proper folding of the protein and occurring at redox potential values approaching apoptotic levels. The dimer binds Ca<sup>2+</sup> only in one EF-hand per monomer, thereby representing a unique state, with decreased α-helicity and thermal stability, increased surface hydrophobicity, and markedly improved inhibitory activity against GRK1 due to 20-fold higher affinity towards the enzyme. Furthermore, dNCS-1 can coordinate zinc and, according to molecular modeling, has an asymmetrical structure and increased conformational flexibility of the subunits, which may underlie their enhanced target-binding properties. In HEK293 cells, dNCS-1 can be reduced by the thioredoxin system, otherwise accumulating as protein aggregates, which are degraded by the proteasome. Interestingly, NCS-1 silencing diminishes the susceptibility of Y79 cancer cells to oxidative stress-induced apoptosis, suggesting that NCS-1 may mediate redox-regulated pathways governing cell death/survival in response to oxidative conditions.
format	article
author	Viktoriia E. Baksheeva Alexey V. Baldin Arthur O. Zalevsky Aliya A. Nazipova Alexey S. Kazakov Vasiliy I. Vladimirov Neonila V. Gorokhovets François Devred Pavel P. Philippov Alexandr V. Bazhin Andrey V. Golovin Andrey A. Zamyatnin Dmitry V. Zinchenko Philipp O. Tsvetkov Sergei E. Permyakov Evgeni Yu. Zernii
author_facet	Viktoriia E. Baksheeva Alexey V. Baldin Arthur O. Zalevsky Aliya A. Nazipova Alexey S. Kazakov Vasiliy I. Vladimirov Neonila V. Gorokhovets François Devred Pavel P. Philippov Alexandr V. Bazhin Andrey V. Golovin Andrey A. Zamyatnin Dmitry V. Zinchenko Philipp O. Tsvetkov Sergei E. Permyakov Evgeni Yu. Zernii
author_sort	Viktoriia E. Baksheeva
title	Disulfide Dimerization of Neuronal Calcium Sensor-1: Implications for Zinc and Redox Signaling
title_short	Disulfide Dimerization of Neuronal Calcium Sensor-1: Implications for Zinc and Redox Signaling
title_full	Disulfide Dimerization of Neuronal Calcium Sensor-1: Implications for Zinc and Redox Signaling
title_fullStr	Disulfide Dimerization of Neuronal Calcium Sensor-1: Implications for Zinc and Redox Signaling
title_full_unstemmed	Disulfide Dimerization of Neuronal Calcium Sensor-1: Implications for Zinc and Redox Signaling
title_sort	disulfide dimerization of neuronal calcium sensor-1: implications for zinc and redox signaling
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/49fd7d64d9f04e46b0edbc351e7d15fc
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Disulfide Dimerization of Neuronal Calcium Sensor-1: Implications for Zinc and Redox Signaling

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