A Strategic Approach for Fluorescence Imaging of Membrane Proteins in a Native-like Environment
Name
nihms-1545063.pdf
Description
Accepted version
Size
1.86 MB
Format
Adobe PDF
Checksum (MD5)
e58447757ce40886a0cd44e8ac14d733
Author(s)
Swiecicki, Jean-Marie
Santana, Jordan Tyler
Imperiali, Barbara
Date Issued
2020
Journal
Cell Chemical Biology
Publisher
Elsevier BV
Version
Author's final manuscript
Abstract
© 2019 Elsevier Ltd Biological membranes are complex barriers in which membrane proteins and thousands of lipidic species participate in structural and functional interactions. Developing a strategic approach that allows uniform labeling of membrane proteins while maintaining a lipidic environment that retains functional interactions is highly desirable for in vitro fluorescence studies. Herein, we focus on complementing current methods by integrating the powerful processes of unnatural amino acid mutagenesis, bioorthogonal labeling, and the detergent-free membrane protein solubilization based on the amphiphilic styrene-maleic acid (SMA) polymer. Importantly, the SMA polymer preserves a thermodynamically stable shell of phospholipids. The approach that we present is both rapid and generalizable providing a population of uniquely labeled membrane proteins in lipid nanoparticles for quantitative fluorescence-based studies.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Physics
Massachusetts Institute of Technology. Department of Chemistry
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs License
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1016/J.CHEMBIOL.2019.11.008
