Two-dimensional crystallization of a membrane protein on a detergent-resistant lipid monolayer.
J Mol Biol
; 308(4): 639-47, 2001 May 11.
Article
em En
| MEDLINE
| ID: mdl-11350166
ABSTRACT
Two-dimensional crystals of a membrane protein, the proton ATPase from plant plasma membranes, have been obtained by a new strategy based on the use of functionalized, fluorinated lipids spread at the air-water interface. Monolayers of the fluorinated lipids are stable even in the presence of high concentrations of various detergents as was established by ellipsometry measurements. A nickel functionalized fluorinated lipid was spread into a monolayer at the air-water interface. The overexpressed His-tagged ATPase solubilized by detergents was added to the subphase. 2D crystals of the membrane protein, embedded in a lipid bilayer, formed as the detergent was removed by adsorption. Electron microscopy indicated that the 2D crystals were single layers with dimensions of 10 microm or more. Image processing yielded a projection map at 9 A resolution, showing three well-separated domains of the membrane-embedded proton ATPase.
Buscar no Google
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Microscopia Crioeletrônica
/
Detergentes
/
Metabolismo dos Lipídeos
/
Proteínas de Membrana
/
Membranas Artificiais
Idioma:
En
Revista:
J Mol Biol
Ano de publicação:
2001
Tipo de documento:
Article
País de afiliação:
França