la9b01288_si_001.pdf (354.38 kB)

Lipid and Protein Transfer between Nanolipoprotein Particles and Supported Lipid Bilayers

Download (354.38 kB)
journal contribution
posted on 06.09.2019, 13:07 by Amanda T. Dang, Wei He, Daniela B. Ivey, Matthew A. Coleman, Tonya L. Kuhl
A nanolipoprotein particle (NLP) is a lipid bilayer disc stabilized by two amphipathic “scaffold” apolipoproteins. It has been most notably utilized as a tool for solubilizing a variety of membrane proteins while preserving structural and functional properties. Transfer of functional proteins from NLPs into model membrane systems such as supported lipid bilayers (SLBs) would enable new opportunities, for example, two-dimensional protein crystallization and studies on protein–protein interactions. This work used fluorescence microscopy and atomic force microscopy to investigate the interaction between NLPs and SLBs. When incubated with SLBs, NLPs were found to spontaneously deliver lipid and protein cargo. The impact of membrane composition on lipid exchange was explored, revealing a positive correlation between the magnitude of lipid transfer and concentration of defects in the target SLB. Incorporation of lipids capable of binding specifically to polyhistidine tags encoded into the apolipoproteins also boosted transfer of NLP cargo. Optimal conditions for lipid and protein delivery from NLPs to SLBs are proposed based on interaction mechanisms.