コンテンツへスキップ
Merck
  • Crystallizing membrane proteins for structure determination using lipidic mesophases.

Crystallizing membrane proteins for structure determination using lipidic mesophases.

Journal of visualized experiments : JoVE (2010-11-30)
Martin Caffrey, Christopher Porter
要旨

A detailed protocol for crystallizing membrane proteins by using lipidic mesophases is described. This method has variously been referred to as the lipidic cubic phase or in meso method. The method has been shown to be quite versatile in that it has been used to solve X-ray crystallographic structures of prokaryotic and eukaryotic proteins, proteins that are monomeric, homo- and hetero-multimeric, chromophore-containing and chromophore-free, and alpha-helical and beta-barrel proteins. Recent successes using in meso crystallization are the human engineered beta2-adrenergic and adenosine A2a G protein-coupled receptors. Protocols are presented for reconstituting the membrane protein into the monoolein-based mesophase, and for setting up crystallizations in the manual mode. Additional steps in the overall process, such as crystal harvesting, are to be addressed in future video articles. The time required to prepare the protein-loaded mesophase and to set up a crystallization plate manually is about one hour.

材料
製品番号
ブランド
製品内容

Avanti
9.7 MAG, Avanti Research - A Croda Brand
Avanti
7.7 MAG, Avanti Research - A Croda Brand 850530O
Avanti
7.9 MAG, Avanti Research - A Croda Brand 850534O
Avanti
8.7 MAG, Avanti Research - A Croda Brand 850532P, powder
Avanti
8.9 MAG, 1-(8Z-heptadecenoyl)-rac-glycerol, powder
Avanti
8.8 MAG, Avanti Research - A Croda Brand 850536O
Avanti
8.6 MAG, Avanti Research - A Croda Brand 850533P, powder