Curved Membrane Mimics for Quantitative Probing of Protein-Membrane Interactions by SPR
Description:
To date the vast majority of biomimetic membrane mimics used to investigate biophysical interactions between lipid membranes and proteins are flat membrane mimics, formed through either supported lipid bilayers (SLBs) or self-assembled monolayers (SAMs). While they have allowed for a great deal of information to be collected, lack of curvature makes these models ineffective for the investigation of curvature sensitive biophysical interactions. To understand the effect of endosomal membrane on membrane interactions and proteins that rely on curvature, such as BAR domain containing proteins, there is a clear need for a reliable platform to generate reproducible and modifiable curved membrane mimics. In this work, we report the development of a technique that can reliably generate curved membrane mimics with the ability to incorporate various cellular membrane components with ease. The curved membrane platform is generated through the use of an underlayer membrane composed of NTA-lipids on a silicated gold substrate. The interaction with his-tagged CT provided the needed orientation to ensure the binding to lipid vesicles containing controlled percentage of gangliosides, including GA1, GM1, GT1b, and GQ1b. Characterization of the curved membrane platform was accomplished with surface plasmon resonance (SPR), fluorescence recovery after photobleaching (FRAP), and nano tracking analysis (NTA). Further investigation of the platform was achieved through Monte Carlo methods to simulate and predict how curvature impacts the distribution of the gangliosides. This platform allows examination of potential monovalent or multivalent interactions with the target proteins, and provides insights for optimizing sensing interfaces toward maximal detection sensitivity.
Speaker: Alexander Malinick - University of California Riverside
Co-Authors
Curved Membrane Mimics for Quantitative Probing of Protein-Membrane Interactions by SPR
Category
2023 Call for Oral Abstracts
Description
Session Number: C51-06
Session Type:
Session Date: Wednesday 3/22/2023
Session Time: 8:30 AM - 11:25 AM
Room Number: 115B
Track: Bioanalytics & Life Sciences
Category: Bioanalytical, Life Sciences, Material Science
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