Functional Status of Neuronal Calcium Sensor-1 Is Modulated by Zinc Binding

Tsvetkov, Philipp O.; Roman, Andrei; Baksheeva, Viktoriia E.; Nazipova, Aliya A.; Shevelyova, Marina P.; Vladimirov, Vasiliy I.; Buyanova, Michelle F.; Zinchenko, D. V.; Zamyatnin, Andrey A.; Devred, François; Golovin, Andrey V.; Permyakov, Sergei E.; Zernii, Evgeni Yu

doi:10.3389/fnmol.2018.00459

Cited by 27 publications

(56 citation statements)

References 103 publications

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“…Zinc can bind not only to a number of prone-toaggregate proteins implicated in neurodegeneration (such as amyloid-β [17], FUS/TLS [18], α-synuclein [19], TDP-43 [20,21], etc.) and favor their aggregation, but also induce aggregation of stable proteins [22].…”

Section: Introductionmentioning

confidence: 99%

Zinc Induces Temperature-Dependent Reversible Self-Assembly of Tau

Roman

Devred

Byrne

et al. 2019

Journal of Molecular Biology

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Tau is an intrinsically disordered microtubule-associated protein that is implicated in several neurodegenerative disorders called tauopathies. In these diseases, Tau is found in the form of intracellular inclusions that consist of aggregated paired helical filaments (PHFs) in neurons. Given the importance of this irreversible PHF formation in neurodegenerative disease, Tau aggregation has been extensively studied. Several different factors, such as mutations or post translational modifications, have been shown to influence the formation of late-stage non-reversible Tau aggregates. It was recently shown that zinc ions accelerated heparin-induced oligomerization of Tau constructs. Indeed, in vitro studies of PHFs have usually been performed in the presence of additional co-factors, such as heparin, in order to accelerate their formation. Using turbidimetry, we investigated the impact of zinc ions on Tau in the absence of heparin and found that zinc is able to induce a temperature-dependent reversible oligomerization of Tau. The obtained oligomers were not amyloid-like and dissociated instantly following zinc chelation or a temperature decrease. Finally, a combination of isothermal titration calorimetry and dynamic light scattering experiments showed zinc binding to a high-affinity binding site and three low-affinity sites on Tau, accompanied by a change in Tau folding. Altogether, our findings stress the importance of zinc in Tau oligomerization. This newly identified Zn-induced oligomerization mechanism may be a part of a pathway different of and concurrent to Tau aggregation cascade leading to PHF formation.

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Section: Introductionmentioning

confidence: 99%

Zinc Induces Temperature-Dependent Reversible Self-Assembly of Tau

Roman

Devred

Byrne

et al. 2019

Journal of Molecular Biology

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“…Competent Escherichia coli cells (strain BL-21(DE3)CodonPlus) were co-transformed with vectors encoding NCS-1 WT and PBB131 plasmid containing the myristoyl-CoA:protein N-myristoyltransferase (NMT) gene from Saccharomyces cerevisiae. Co-expression of NCS-1 variants with NMT and subsequent purification of the myristoylated proteins from bacterial lysate was carried out as previously described for NCS-1 WT [31]. Myristoylation levels of the produced proteins were determined by analytical high-performance liquid chromatography (HPLC) using a reversed-phase column (Phenomenex Luna C18(2)) [32].…”

Section: Purification and Characterization Of Myristoylated And Unmyrmentioning

confidence: 99%

“…NCS-1 binding to urea-washed membranes was performed by equilibrium centrifugation assay, as described in [31,36] with modifications. Briefly, 5 µL of the suspension of photoreceptor or hippocampal membranes were mixed with 40 µM NCS-1 in 20 mM Tris-HCl buffer (pH 8.0), 150 mM NaCl, 1 mM DTT, 20 mM MgCl 2 and either 2 mM CaCl 2 or 2 mM EGTA in the total volume of 50 µL.…”

Section: Membrane Binding Assaymentioning

confidence: 99%

Membrane Binding of Neuronal Calcium Sensor-1: Highly Specific Interaction with Phosphatidylinositol-3-Phosphate

et al. 2020

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Neuronal calcium sensors are a family of N-terminally myristoylated membrane-binding proteins possessing a different intracellular localization and thereby targeting unique signaling partner(s). Apart from the myristoyl group, the membrane attachment of these proteins may be modulated by their N-terminal positively charged residues responsible for specific recognition of the membrane components. Here, we examined the interaction of neuronal calcium sensor-1 (NCS-1) with natural membranes of different lipid composition as well as individual phospholipids in form of multilamellar liposomes or immobilized monolayers and characterized the role of myristoyl group and N-terminal lysine residues in membrane binding and phospholipid preference of the protein. NCS-1 binds to photoreceptor and hippocampal membranes in a Ca2+-independent manner and the binding is attenuated in the absence of myristoyl group. Meanwhile, the interaction with photoreceptor membranes is less dependent on myristoylation and more sensitive to replacement of K3, K7, and/or K9 of NCS-1 by glutamic acid, reflecting affinity of the protein to negatively charged phospholipids. Consistently, among the major phospholipids, NCS-1 preferentially interacts with phosphatidylserine and phosphatidylinositol with micromolar affinity and the interaction with the former is inhibited upon mutating of N-terminal lysines of the protein. Remarkably, NCS-1 demonstrates pronounced specific binding to phosphoinositides with high preference for phosphatidylinositol-3-phosphate. The binding does not depend on myristoylation and, unexpectedly, is not sensitive to the charge inversion mutations. Instead, phosphatidylinositol-3-phosphate can be recognized by a specific site located in the N-terminal region of the protein. These data provide important novel insights into the general mechanism of membrane binding of NCS-1 and its targeting to specific phospholipids ensuring involvement of the protein in phosphoinositide-regulated signaling pathways.

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“…Due to specific structural features, such as different organization of Ca 2+ -binding loops and the presence of specific regulatory elements (for instance, C-terminal segment [27][28][29]), each NCS binds calcium within its own narrow range of the cation concentration, which allows these proteins to respond to different Ca 2+ -signals and play the role of Ca 2+ -buffer in cells [30]. Alongside calcium and magnesium, NCSs are capable of binding zinc, which is suggested to be important for their normal and pathological activity [31,32].…”

Section: Introductionmentioning

confidence: 99%

A Novel Approach to Bacterial Expression and Purification of Myristoylated Forms of Neuronal Calcium Sensor Proteins

et al. 2020

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N-terminal myristoylation is a common co-and post-translational modification of numerous eukaryotic and viral proteins, which affects their interaction with lipids and partner proteins, thereby modulating various cellular processes. Among those are neuronal calcium sensor (NCS) proteins, mediating transduction of calcium signals in a wide range of regulatory cascades, including reception, neurotransmission, neuronal growth and survival. The details of NCSs functioning are of special interest due to their involvement in the progression of ophthalmological and neurodegenerative diseases and their role in cancer. The well-established procedures for preparation of native-like myristoylated forms of recombinant NCSs via their bacterial co-expression with N-myristoyl transferase from Saccharomyces cerevisiae often yield a mixture of the myristoylated and non-myristoylated forms. Here, we report a novel approach to preparation of several NCSs, including recoverin, GCAP1, GCAP2, neurocalcin δ and NCS-1, ensuring their nearly complete N-myristoylation. The optimized bacterial expression and myristoylation of the NCSs is followed by a set of procedures for separation of their myristoylated and non-myristoylated forms using a combination of hydrophobic interaction chromatography steps. We demonstrate that the refolded and further purified myristoylated NCS-1 maintains its Са2+-binding ability and stability of tertiary structure. The developed approach is generally suited for preparation of other myristoylated proteins.

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Functional Status of Neuronal Calcium Sensor-1 Is Modulated by Zinc Binding

Cited by 27 publications

References 103 publications

Zinc Induces Temperature-Dependent Reversible Self-Assembly of Tau

Zinc Induces Temperature-Dependent Reversible Self-Assembly of Tau

Membrane Binding of Neuronal Calcium Sensor-1: Highly Specific Interaction with Phosphatidylinositol-3-Phosphate

A Novel Approach to Bacterial Expression and Purification of Myristoylated Forms of Neuronal Calcium Sensor Proteins

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