Interaction of membrane proteins and lipids with solubilizing detergents.
Detergents are indispensable in the isolation of integral membrane proteins from biological membranes to study their intrinsic structural and functional properties. Solubilization involves a number of intermediary states that can be studied by a variety of physicochemical and kinetic methods; it usually starts by destabilization of the lipid component of the membranes, a process that is accompanied by a transition of detergent binding by the membrane from a noncooperative to a cooperative interaction already below the critical micellar concentration (CMC). This leads to the formation of membrane fragments of proteins and lipids with detergent-shielded edges. In the final stage of solubilization membrane proteins are present as protomers, with the membrane inserted sectors covered by detergent. We consider in detail the nature of this interaction and conclude that in general binding as a monolayer ring, rather than as a micelle, is the most probable mechanism. This mode of interaction is supported by neutron diffraction investigations on the disposition of detergent in 3-D crystals of membrane proteins. Finally, we briefly discuss the use of techniques such as analytical ultracentrifugation, size exclusion chromatography, and mass spectrometry relevant for the structural investigation of detergent solubilized membrane proteins.
le Maire
M
M
Unite de recherche Associée 2096 (Centre National de la Recherche Scientifique et Commissariat a l'Energie Atomique), Cedex, France. lemairem@dsvidf.cea.fr
Champeil
P
P
Moller
J V
JV
D003461:Crystallography; D003902:Detergents / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry; D008563:Membrane Lipids / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry*; D008823:Micelles; D015394:Molecular Structure; D011092:Polyethylene Glycols / Q000737:chemistry; D011487:Protein Conformation; D012542:Scattering, Radiation; D012995:Solubility; D013501:Surface-Active Agents / Q000737:chemistry*
D003902:Detergents; D008051:Lipid Bilayers; D008563:Membrane Lipids; D008565:Membrane Proteins; D008823:Micelles; D013501:Surface-Active Agents; D011092:Polyethylene Glycols