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Characterization of neurocalcin delta membrane binding by biophysical methods
Emmanuelle Hoareau1, Nicolas Belley1, Kristina Klinker1
1CUO-Recherche, Hôpital du Saint-Sacrement, Centre de recherche du CHU de Québec and Département d'ophtalmologie, Faculté de médecine, Université Laval, Québec, Québec, G3K 1A3, Canada.
Colloids and Surfaces. B, Biointerfaces
|November 24, 2018
Summary
Neurocalcin delta (NCALD), a neuronal calcium sensor, binds membranes via a calcium-myristoyl switch. Its interaction is influenced by lipid composition and calcium levels, potentially explaining its role in Alzheimer's disease.
Area of Science:
- Neuroscience
- Biochemistry
- Cell Biology
Background:
- Neurocalcin delta (NCALD) is a neuronal calcium sensor protein family member expressed in retinal ganglion and amacrine cells.
- NCALD possesses EF-hand motifs, and calcium binding triggers a conformational change involving its N-terminal myristoyl group, termed the calcium-myristoyl switch.
- The precise mechanisms governing NCALD's membrane interaction remain incompletely understood.
Purpose of the Study:
- To investigate the membrane binding behavior of NCALD.
- To elucidate the influence of lipid composition, calcium, and the myristoyl group on NCALD-membrane interactions.
- To determine key binding parameters like Maximum Insertion Pressure (MIP) and synergy.
Main Methods:
- Utilized lipid monolayers as a model membrane system to study NCALD binding.
- Injected NCALD underneath various lipid monolayers to measure binding parameters.
- Employed polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) and fluorescence microscopy for detailed analysis.
Main Results:
- NCALD exhibited higher Maximum Insertion Pressure (MIP) with saturated phospholipids featuring phosphoethanolamine head groups.
- Polarization modulation infrared reflection absorption spectroscopy confirmed these findings.
- Fluorescence microscopy revealed NCALD preferentially interacts with phospholipids in the liquid-condensed phase, characteristic of membrane microdomains.
Conclusions:
- NCALD's membrane binding is significantly influenced by specific lipid compositions and calcium presence.
- The preferential interaction with liquid-condensed phase phospholipids suggests a role in membrane microdomains.
- Altered lipid composition in Alzheimer's disease microdomains may impact NCALD expression levels, warranting further investigation.
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