Summary of 125-Gessesse-CX3CR1nanodisc

SSBD:database URL
Transmission electron images of homogeneity of CX3CR1-nanodisc complex in a reconstituted cell-free synthesis system
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Image data based on Experiment
Number of Datasets
1 ( Image datasets: 1, Quantitative data datasets: 0 )
Size of Datasets
412.7 MB ( Image datasets: 412.7 MB, Quantitative data datasets: 0 bytes )


protein analysis
Molecular Function (MF)
Biological Process (BP)
protein transport within lipid bilayer
Cellular Component (CC)
membrane protein complex, G protein coupled receptor complex
Biological Imaging Method
XYZ Scale
XY: 1 micrometer/pixel, Z: 1 micrometer/slice
T scale

Image Acquisition
Experiment type
Microscope type
Acquisition mode
Contrast method
Microscope model
Hitachi H-7000 electron microscopy
Detector model
Objective model
Filter set

Related paper(s)

Belay Gessesse, Takashi Nagaike, Koji Nagata, Yoshihiro Shimizu, Takuya Ueda (2018) G-Protein Coupled Receptor Protein Synthesis on a Lipid Bilayer Using a Reconstituted Cell-Free Protein Synthesis System., Life (Basel, Switzerland), Volume 8, Number 4

Published in 2018 Nov 2 (Electronic publication in Nov. 2, 2018, midnight )

(Abstract) Membrane proteins are important drug targets which play a pivotal role in various cellular activities. However, unlike cytosolic proteins, most of them are difficult-to-express proteins. In this study, to synthesize and produce sufficient quantities of membrane proteins for functional and structural analysis, we used a bottom-up approach in a reconstituted cell-free synthesis system, the PURE system, supplemented with artificial lipid mimetics or micelles. Membrane proteins were synthesized by the cell-free system and integrated into lipid bilayers co-translationally. Membrane proteins such as the G-protein coupled receptors were expressed in the PURE system and a productivity ranging from 0.04 to 0.1 mg per mL of reaction was achieved with a correct secondary structure as predicted by circular dichroism spectrum. In addition, a ligand binding constant of 27.8 nM in lipid nanodisc and 39.4 nM in micelle was obtained by surface plasmon resonance and the membrane protein localization was confirmed by confocal microscopy in giant unilamellar vesicles. We found that our method is a promising approach to study the different classes of membrane proteins in their native-like artificial lipid bilayer environment for functional and structural studies.

Yoshihiro Shimizu , RIKEN , Center for Biosystems Dynamics Research , Laboratory for Cell-Free Protein Synthesis
Belay Gessesse, Takashi Nagaike, Koji Nagata, Yoshihiro Shimizu, Takuya Ueda

Dataset List of 125-Gessesse-CX3CR1nanodisc

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# 4898
Dataset Kind Image data
Dataset Size 412.7 MB
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