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Oligomeric organization of membrane proteins from native membranes at nanoscale spatial and single-molecule resolution
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Oligomeric organization of membrane proteins from native membranes at nanoscale spatial and single-molecule resolution
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Journal Article
Oligomeric organization of membrane proteins from native membranes at nanoscale spatial and single-molecule resolution
2024
Overview
The oligomeric organization of membrane proteins in native cell membranes is a critical regulator of their function. High-resolution quantitative measurements of oligomeric assemblies and how they change under different conditions are indispensable to understanding membrane protein biology. We report Native-nanoBleach, a total internal reflection fluorescence microscopy-based single-molecule photobleaching step analysis technique to determine the oligomeric distribution of membrane proteins directly from native membranes at an effective spatial resolution of ~10 nm. We achieved this by capturing target membrane proteins in native nanodiscs with their proximal native membrane environment using amphipathic copolymers. We applied Native-nanoBleach to quantify the oligomerization status of structurally and functionally diverse membrane proteins, including a receptor tyrosine kinase (TrkA) and a small GTPase (KRas) under growth-factor binding and oncogenic mutations, respectively. Our data suggest that Native-nanoBleach provides a sensitive, single-molecule platform to quantify membrane protein oligomeric distributions in native membranes under physiologically and clinically relevant conditions.
Native-nanoBleach, a single-molecule imaging technique with a spatial resolution of ~10 nm, quantifies the oligomeric distribution of membrane proteins directly from native membranes at endogenous expression levels with their proximal native membrane environment using amphipathic copolymer nanodiscs.
Publisher
Nature Publishing Group UK,Nature Publishing Group
