Oligomeric organization of membrane proteins from native membranes at nanoscale spatial and single-molecule resolution.
Nat Nanotechnol
; 19(1): 85-94, 2024 Jan.
Article
em En
| MEDLINE
| ID: mdl-38012273
ABSTRACT
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.
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Base de dados:
MEDLINE
Assunto principal:
Proteínas de Membrana
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article