RT Journal Article
SR Electronic
T1 Determination of oligomeric organization of membrane proteins from native membranes at nanoscale-spatial and single-molecule resolution
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2023.02.19.529138
DO 10.1101/2023.02.19.529138
A1 Gerard Walker
A1 Caroline Brown
A1 Xiangyu Ge
A1 Shailesh Kumar
A1 Mandar D. Muzumdar
A1 Kallol Gupta
A1 Moitrayee Bhattacharyya
YR 2023
UL http://biorxiv.org/content/early/2023/02/21/2023.02.19.529138.abstract
AB 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 the understanding of membrane protein biology. We report a single-molecule imaging technique (Native-nanoBleach) 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 established this method using structurally and functionally diverse membrane proteins with well-established stoichiometries. We then applied Native-nanoBleach to quantify the oligomerization status of a receptor tyrosine kinase (TrkA) and a small GTPase (KRas) under conditions of growth-factor binding or oncogenic mutations, respectively. Native-nanoBleach provides a sensitive, single-molecule platform to quantify membrane protein oligomeric distributions in native membranes at an unprecedented spatial resolution.Competing Interest StatementThe authors have declared no competing interest.