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CaF2 nanoparticles as surface carriers of GCAP1, a calcium sensor protein involved in retinal dystrophies

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Abstract

CaF2-based nanoparticles (NP) are promising biocompatible tools for nanomedicine applications. The structure of the NP crystal lattice allows for specific interactions with Ca2+-binding proteins through their EF-hand cation binding motifs. Here we investigated the interaction of 23 nm citrate-coated CaF2 NP with a calcium sensor protein GCAP1 that is normally expressed in photoreceptor cells and involved in the regulation of the early steps of vision. Protein–NP interactions were thoroughly investigated for the wild type (WT) GCAP1 as well as for a variant carrying the Asp 100 to Glu mutation (D100E), which prevents the binding of Ca2+ to the highest affinity site and is linked to cone dystrophy. Circular dichroism and fluorescence spectroscopy showed that protein structure and Ca2+-sensing capability are conserved for both variants upon interaction with the NP surface, although the interaction mode depends on the specific occupation of Ca2+-binding sites. NP binding stabilizes the structure of the bound GCAP1 and occurs with nanomolar affinity, as probed by isothermal titration calorimetry. Surface plasmon resonance revealed a fully reversible binding compatible with physiologically relevant kinetics of protein release whereas biochemical assays indicated a residual capability for NP-dissociated GCAP1 to regulate the target retinal guanylate cyclase. Our study constitutes a proof of concept that CaF2 NP could be optimized to serve as biologically compatible carriers of high amounts of functional GCAP1 in photoreceptors affected by retinal dystrophies.

Graphical abstract: CaF2 nanoparticles as surface carriers of GCAP1, a calcium sensor protein involved in retinal dystrophies

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Publication details

The article was received on 09 May 2017, accepted on 17 Jul 2017 and first published on 08 Aug 2017


Article type: Paper
DOI: 10.1039/C7NR03288A
Citation: Nanoscale, 2017, Advance Article
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    CaF2 nanoparticles as surface carriers of GCAP1, a calcium sensor protein involved in retinal dystrophies

    V. Marino, A. Borsatto, F. Vocke, K. Koch and D. Dell'Orco, Nanoscale, 2017, Advance Article , DOI: 10.1039/C7NR03288A

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