A high-throughput differential filtration assay to screen and select detergents for membrane proteins.
Anal Biochem
; 407(1): 1-11, 2010 Dec 01.
Article
em En
| MEDLINE
| ID: mdl-20667442
ABSTRACT
Structural studies on integral membrane proteins are routinely performed on protein-detergent complexes (PDCs) consisting of purified protein solubilized in a particular detergent. Of all the membrane protein crystal structures solved to date, a subset of only four detergents has been used in more than half of these structures. Unfortunately, many membrane proteins are not well behaved in these four detergents and/or fail to yield well-diffracting crystals. Identification of detergents that maintain the solubility and stability of a membrane protein is a critical step and can be a lengthy and "protein-expensive" process. We have developed an assay that characterizes the stability and size of membrane proteins exchanged into a panel of 94 commercially available and chemically diverse detergents. This differential filtration assay (DFA), using a set of filtered microplates, requires sub-milligram quantities of purified protein and small quantities of detergents and other reagents and is performed in its entirety in several hours.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Detergentes
/
Filtração
/
Proteínas de Membrana
Idioma:
En
Ano de publicação:
2010
Tipo de documento:
Article