Overview of electron crystallography of membrane proteins: crystallization and screening strategies using negative stain electron microscopy.
Curr Protoc Protein Sci
; Chapter 17: Unit17.15, 2013.
Article
en En
| MEDLINE
| ID: mdl-23546618
ABSTRACT
Electron cryomicroscopy, or cryoEM, is an emerging technique for studying the three-dimensional structures of proteins and large macromolecular machines. Electron crystallography is a branch of cryoEM in which structures of proteins can be studied at resolutions that rival those achieved by X-ray crystallography. Electron crystallography employs two-dimensional crystals of a membrane protein embedded within a lipid bilayer. The key to a successful electron crystallographic experiment is the crystallization, or reconstitution, of the protein of interest. This unit describes ways in which protein can be expressed, purified, and reconstituted into well-ordered two-dimensional crystals. A protocol is also provided for negative stain electron microscopy as a tool for screening crystallization trials. When large and well-ordered crystals are obtained, the structures of both protein and its surrounding membrane can be determined to atomic resolution.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Coloración Negativa
/
Microscopía por Crioelectrón
/
Cristalografía
/
Proteínas de la Membrana
Tipo de estudio:
Diagnostic_studies
/
Screening_studies
Idioma:
En
Revista:
Curr Protoc Protein Sci
Año:
2013
Tipo del documento:
Article
País de afiliación:
Estados Unidos