Membrane proteins: functional and structural studies using reconstituted proteoliposomes and 2-D crystals.
Reconstitution of membrane proteins into lipid bilayers is a powerful tool to analyze functional as well as structural areas of membrane protein research. First, the proper incorporation of a purified membrane protein into closed lipid vesicles, to produce proteoliposomes, allows the investigation of transport and/or catalytic properties of any membrane protein without interference by other membrane components. Second, the incorporation of a large amount of membrane proteins into lipid bilayers to grow crystals confined to two dimensions has recently opened a new way to solve their structure at high resolution using electron crystallography. However, reconstitution of membrane proteins into functional proteoliposomes or 2-D crystallization has been an empirical domain, which has been viewed for a long time more like "black magic" than science. Nevertheless, in the last ten years, important progress has been made in acquiring knowledge of lipid-protein-detergent interactions and has permitted to build upon a set of basic principles that has limited the empirical approach of reconstitution experiments. Reconstitution strategies have been improved and new strategies have been developed, facilitating the success rate of proteoliposome formation and 2-D crystallization. This review deals with the various strategies available to obtain proteoliposomes and 2-D crystals from detergent-solubilized proteins. It gives an overview of the methods that have been applied, which may be of help for reconstituting more proteins into lipid bilayers in a form suitable for functional studies at the molecular level and for high-resolution structural analysis.
Rigaud
J-L
JL
Institut Curie, UMR-CNRS 168 and LRC-CEA 8, Paris, France. rigaud@curie.fr
D003460:Crystallization; D018360:Crystallography, X-Ray; D003902:Detergents; D008081:Liposomes / Q000737:chemistry*; D008563:Membrane Lipids / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry* / Q000502:physiology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D011487:Protein Conformation; D011510:Proteolipids / Q000737:chemistry* / Q000648:ultrastructure
D003902:Detergents; D008081:Liposomes; D008563:Membrane Lipids; D008565:Membrane Proteins; D011510:Proteolipids; C020485:proteoliposomes