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Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects
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Kinetically Stable Triglyceride-Based Nanodroplets and Their Interactions with Lipid-Specific Proteins
Valerija Vezočnik1, Vesna Hodnik1, Simona Sitar2
1Department of Biology, Biotechnical Faculty , University of Ljubljana , Jamnikarjeva 101 , Ljubljana 1000 , Slovenia.
Langmuir : the ACS Journal of Surfaces and Colloids
|July 10, 2018
Summary
Researchers developed stable lipid nanodroplets using sphingomyelin, cholesterol, and trioleoylglycerol. These nanoparticles mimic natural lipid droplets and protein interactions with cell membranes.
Area of Science:
- Biophysics
- Nanotechnology
- Biochemistry
Background:
- Understanding protein-lipid interactions is crucial for physiology, pathology, and nanotechnology.
- Cellular droplets necessitate simplified biomimetic lipid models due to in vivo complexity.
Purpose of the Study:
- To present a protocol for preparing kinetically stable nanoemulsions with sphingomyelin (SM), cholesterol (Chol), and trioleoylglycerol (TOG) nanodroplets.
- To establish SM/Chol/TOG nanoparticles as experimental models for monolayer membranes, imitating natural lipid droplets.
Main Methods:
- Modified reverse phase evaporation method combined with ultrasonication for stable SM/Chol-coated monodisperse lipid nanodroplets.
- Characterization of lipid nanodroplets (composition, ζ-potential, size, shape, stability) and comparison with large unilamellar vesicles.
- Investigation of interfacial water molecule order and protein interactions (equinatoxin II, perfringolysin O) using second harmonic scattering.
Main Results:
- Stable SM/Chol/TOG nanodroplets were successfully prepared at various molar ratios.
- Nanodroplets exhibited surface characteristics and protein binding similar to lipid bilayers and Langmuir monolayers.
- The orientational order of interfacial water molecules was investigated using second harmonic scattering.
Conclusions:
- SM/Chol/TOG nanoparticles serve as effective biomimetic models for monolayer membranes.
- These nanoparticles can imitate natural lipid droplets and facilitate studies on protein-lipid interactions.
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