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Glucose-based amphiphilic telomers designed to keep membrane proteins soluble in aqueous solutions: synthesis and physicochemical characterization.

Langmuir : the ACS journal of surfaces and colloids (2008-11-05)
K Shivaji Sharma, Grégory Durand, Fabrice Giusti, Blandine Olivier, Anne-Sylvie Fabiano, Paola Bazzacco, Tassadite Dahmane, Christine Ebel, Jean-Luc Popot, Bernard Pucci
ABSTRACT

A novel class of nonionic amphipols (NAPols) designed to handle membrane proteins in aqueous solutions has been synthesized, and its solution properties have been examined. These were synthesized through free radical cotelomerization of glucose-based hydrophilic and amphiphilic monomers derived from tris(hydroxymethyl)acrylamidomethane using azobisisobutyronitrile as the initiator and thiol as the transfer agent. The molecular weight and the hydrophilic/lipophilic balance of the cotelomers were modulated by varying the thiol/monomers and the hydrophilic monomer/amphiphilic monomer ratios, respectively, and were characterized by 'H NMR, UV, gel permeation chromatography, and Fourier transform infrared spectroscopy. Their physicochemical properties in aqueous solution were studied by dynamic light scattering, aqueous size-exclusion chromatography, analytical ultracentrifugation, and surface-tension measurements. NAPols are highly soluble in water and form, within a large concentration range, well-defined supramolecular assemblies with a diameter of approximately 6-7 nm, a narrow particle size distribution, and an average molecular weight close to 50 x 10(3) g x mol(-1). Varying the hydrophilic/amphiphilic monomer ratio of NAPols in the range of 3.0-4.9, the degree of polymerization in the range of 51-78, and the resulting average molar mass in the range of 20-29 x 10(3) g x mol(-1) has little incidence on their solution properties. Glucose-based NAPols efficiently kept soluble in aqueous solutions two test membrane proteins: bacteriorhodopsin and the transmembrane domain of Escherichia coli's outer membrane protein A.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
2,2′-Azobis(2-methylpropionitrile), 98%
Sigma-Aldrich
2,2′-Azobis(2-methylpropionitrile) solution, 0.2 M in toluene
Sigma-Aldrich
Azobisisobutyronitrile, 12 wt. % in acetone
Sigma-Aldrich
2,2′-Azobis(2-methylpropionitrile), recrystallized from methanol, 99%

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