RRC ID 54329
Author Haruyama T, Sugano Y, Kodera N, Uchihashi T, Ando T, Tanaka Y, Konno H, Tsukazaki T.
Title Single-Unit Imaging of Membrane Protein-Embedded Nanodiscs from Two Oriented Sides by High-Speed Atomic Force Microscopy.
Journal Structure
Abstract Membrane proteins play important roles in various cellular functions. To analyze membrane proteins, nanodisc technology using membrane scaffold proteins allows single membrane protein units to be embedded into the lipid bilayer disc without detergents. Recent advancements in high-speed atomic force microscopy (HS-AFM) have enabled us to monitor the real-time dynamics of proteins in solution at the nanometer scale. In this study, we report HS-AFM imaging of membrane proteins reconstituted into nanodiscs using two membrane protein complexes, SecYEG complex and MgtE dimer. The observed images showed single particles of membrane protein-embedded nanodiscs in an end-up orientation whereby the membrane was fixed parallel to the supporting solid surface and in a side-on orientation whereby the membrane plane was vertically fixed to the solid surface, enabling the elucidation of domain fluctuations in membrane proteins. This technique provides a basic method for the high-resolution imaging of single membrane proteins by HS-AFM.
Volume 27(1)
Pages 152-160.e3
Published 2019-1-2
DOI 10.1016/j.str.2018.09.005
PII S0969-2126(18)30331-9
PMID 30318467
MeSH Antiporters / chemistry* Bacterial Proteins / chemistry* Lipid Bilayers / chemistry Microscopy, Atomic Force / methods Nanoparticles / chemistry* SEC Translocation Channels / chemistry* Single Molecule Imaging / methods
IF 4.862
Times Cited 4
DNA material Genomic DNA of Thermus thermophilus HB8 JCM 10941 T (JGD05989)