title: Nanodevice-induced conformational and functional changes in a prototypical calcium sensor protein.
abstract: Calcium (Ca(2+)) plays a major role in a variety of cellular processes. Fine changes in its concentration are detected by calcium sensor proteins, which adopt specific conformations to regulate their molecular targets. Here, two distinct nanodevices were probed as biocompatible carriers of Ca(2+)-sensors and the structural and functional effects of protein-nanodevice interactions were investigated. The prototypical Ca(2+)-sensor recoverin (Rec) was incubated with 20-25 nm CaF2 nanoparticles (NPs) and 70-80 nm liposomes with lipid composition similar to that found in photoreceptor cells. Circular dichroism and fluorescence spectroscopy were used to characterize changes in the protein secondary and tertiary structure and in thermal stability upon interaction with the nanodevice, both in the presence and in the absence of free Ca(2+). Variations in the hydrodynamic diameter of the complex were measured by dynamic light scattering and the residual capability of the protein to act as a Ca(2+)-sensor in the presence of NPs was estimated spectroscopically. The conformation, thermal stability and Ca(2+)-sensing capability of Rec were all significantly affected by the presence of NPs, while liposomes did not significantly perturb Rec conformation and function, allowing reversible binding. NP-bound Rec maintained an all-helical fold but showed lower thermal stability and high cooperativity of unfolding. Our analysis can be proficiently used to validate the biocompatibility of other nanodevices intended for biomedical applications involving Ca(2+)-sensors.
authors.affiliation: Department of Life Sciences and Reproduction, Section of Biological Chemistry, University of Verona, Verona, Italy. daniele.dellorco@univr.it.
authors.forename: Valerio
authors.initials: V
authors.lastname: Marino
authors.forename: Alessandra
authors.initials: A
authors.lastname: Astegno
authors.forename: Marco
authors.initials: M
authors.lastname: Pedroni
authors.forename: Fabio
authors.initials: F
authors.lastname: Piccinelli
authors.forename: Daniele
authors.initials: D
authors.lastname: Dell'Orco
mesh_terms: D002124:Calcium Fluoride / Q000737:chemistry*; D002942:Circular Dichroism; D008027:Light; D008081:Liposomes / Q000737:chemistry; D053758:Nanoparticles / Q000737:chemistry*; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D051603:Recoverin / Q000737:chemistry* / Q000378:metabolism; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence
chemical_list: D008081:Liposomes; D051603:Recoverin; D002124:Calcium Fluoride