Using supported bilayers to study the spatiotemporal organization of membrane-bound proteins.
Methods Cell Biol
; 128: 223-241, 2015.
Article
in En
| MEDLINE
| ID: mdl-25997350
ABSTRACT
Cell division in prokaryotes and eukaryotes is commonly initiated by the well-controlled binding of proteins to the cytoplasmic side of the cell membrane. However, a precise characterization of the spatiotemporal dynamics of membrane-bound proteins is often difficult to achieve in vivo. Here, we present protocols for the use of supported lipid bilayers to rebuild the cytokinetic machineries of cells with greatly different dimensions the bacterium Escherichia coli and eggs of the vertebrate Xenopus laevis. Combined with total internal reflection fluorescence microscopy, these experimental setups allow for precise quantitative analyses of membrane-bound proteins. The protocols described to obtain glass-supported membranes from bacterial and vertebrate lipids can be used as starting points for other reconstitution experiments. We believe that similar biochemical assays will be instrumental to study the biochemistry and biophysics underlying a variety of complex cellular tasks, such as signaling, vesicle trafficking, and cell motility.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Xenopus laevis
/
Cell Membrane
/
Escherichia coli
/
Lipid Bilayers
/
Membrane Proteins
Limits:
Animals
Language:
En
Journal:
Methods Cell Biol
Year:
2015
Document type:
Article
Affiliation country:
United States