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Detergent removal by non-polar polystyrene beads (1998)

Rigaud, J.-L. ; Levy, Daniel ; Mosser, Gervaise ; [et al.]

Springer

European biophysics journal 27 (1998), S. 305-319

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ISSN: 1432-1017

Keywords: Key words Bio-Beads ; Detergents ; Liposomes ; Proteoliposomes ; Reconstitution ; 2 D crystallization ; Membrane proteins

Source: Springer Online Journal Archives 1860-2000

Topics: Biology , Physics

Notes: Abstract Detergent removal from lipid-protein-detergent micellar solutions is the most successful strategy for reconstitution of integral membrane proteins into proteoliposomes or into two-dimensional crystals. This review establishes the potential of polystyrene beads as a simple alternative to other conventional detergent removing strategies such as dialysis, gel chromatography and dilution. Kinetics and equilibrium aspects of removal of different detergents by hydrophobic adsorption onto polystyrene beads have been systematically investigated. A mechanism of adsorption onto polystyrene beads is proposed and provide useful information about the use of these beads in reconstitution experiments. The usefulness of this detergent removal strategy to produce quasi-ideal proteoliposomes is evaluated by considering the morphology and the size of the reconstituted vesicles, the homogeneity in size and protein distribution, the final protein orientation and the permeability of resulting proteoliposomes. Finally, the advantages of detergent removal by polystyrene beads as an alternative to conventional dialysis in two-dimensional crystallization trials are evaluated through review of recent structural reconstitution studies.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/s002490050138

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Structural analysis of two-dimensional arrays of cholera toxin b-subunit (1991)

Mosser, Gervaise ; Brisson, Alain

New York, NY : Wiley-Blackwell

Journal of Electron Microscopy Technique 18 (1991), S. 387-394

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ISSN: 0741-0581

Keywords: Two-dimensional crystals ; Cholera toxin ; Electron crystallographic analysis ; Oligomeric structure ; Negative staining ; Life and Medical Sciences ; Cell & Developmental Biology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Natural Sciences in General

Notes: Two-dimensional arraysThe term array is used to describe a regular arrangement of motifs without implying the existence of translational symmetry elements between motifs, by distinction with a crystal. of cholera toxin B-subunit (CTB) have been obtained by specific interaction with lipid films, as described by Ludwig et al. (1986). The relationship between two types of array, of either rectangular or hexagonal geometry, was analyzed using crystallographic methods of electron image analysis. Our results showed that the type of array obtained was highly dependent on the negative stain used and that both arrays presented related lattice parameters, indicating that they originated from a common unstained structure. Image analysis of hexagonal arrays at 17 Å resolution revealed variable CTB projected structures, ranging from annularly symmetric particles to highly asymmetric particles, very distinct from the pentameric structure resolved from rectangular crystals. The present data suggest that hexagonal arrays result from an imperfect staining of CTB rectangular crystals. The staining distortion is such that the stain layer does not match faithfully the pentameric protein distribution whereas the regular organization of the specimen is maintained.

Additional Material: 5 Ill.