title: Modeling detergent organization around aquaporin-0 using small-angle X-ray scattering.
abstract: Solubilization of integral membrane proteins in aqueous solutions requires the presence of amphiphilic molecules like detergents. The transmembrane region of the proteins is then surrounded by a corona formed by these molecules, ensuring a hydrophilic outer surface. The presence of this corona has strongly hampered structural studies of solubilized membrane proteins by small-angle X-ray scattering (SAXS), a technique frequently used to monitor conformational changes of soluble proteins. Through the online combination of size exclusion chromatography, SAXS, and refractometry, we have determined a precise geometrical model of the n-dodecyl β-d-maltopyranoside corona surrounding aquaporin-0, the most abundant membrane protein of the eye lens. The SAXS data were well-fitted by a detergent corona shaped in an elliptical toroid around the crystal structure of the protein, similar to the elliptical shape recently reported for nanodiscs (Skar-Gislinge et al. J. Am. Chem. Soc. 2010, 132, 13713-13722). The torus thickness determined from the curve-fitting protocol is in excellent agreement with the thickness of a lipid bilayer, while the number of detergent molecules deduced from the volume of the torus compares well with those obtained on the same sample from refractometry and mass analysis based on SAXS forward scattering. For the first time, the partial specific volume of the detergent surrounding a protein was measured. The present protocol is a crucial step toward future conformational studies of membrane proteins in solution.
authors.affiliation: Institut Curie, Centre de Recherche, CNRS UMR168, Université Pierre et Marie Curie, F-75248 Paris Cedex, France.
authors.forename: Alice
authors.initials: A
authors.lastname: Berthaud
authors.forename: John
authors.initials: J
authors.lastname: Manzi
authors.forename: Javier
authors.initials: J
authors.lastname: Pérez
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Mangenot
mesh_terms: D000818:Animals; D020346:Aquaporins / Q000737:chemistry*; D003902:Detergents / Q000737:chemistry*; D005136:Eye Proteins / Q000737:chemistry*; D007908:Lens, Crystalline / Q000737:chemistry*; D008320:Maltose / Q000031:analogs & derivatives* / Q000737:chemistry; D008958:Models, Molecular; D053838:Scattering, Small Angle; D012756:Sheep; D012995:Solubility; D014961:X-Ray Diffraction
chemical_list: D020346:Aquaporins; D003902:Detergents; D005136:Eye Proteins; C030328:aquaporin 0; C109906:n-lauryl beta-d-maltopyranoside; D008320:Maltose