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Hydrogenated diglucose detergents for membrane-protein extraction and stabilization
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Edité par CCSD ; American Chemical Society -
International audience. We report herein the design and synthesis of a novel series of alkyl glycosides detergents consisting of a nonionic polar headgroup that comprises two glucose moieties in a branched arrangement (DG), onto which octane-, decane- and dodecanethiols were grafted leading to ODG, DDG, and DDDG detergents. Micellization in aqueous solution was studied by isothermal titration calorimetry (ITC), 1H NMR spectroscopy, and surface tensiometry (SFT). Critical micellar concentration values were found to decrease by a factor of ~10 for each pair of methylene groups added to the alkyl chain, ranging from ~0.05 mM to 9 mM for DDDG and ODG, respectively. Dynamic light scattering (DLS) and analytical ultracentrifugation sedimentation velocity (AUC-SV) experiments were used to investigate the size and the composition of the micellar aggregates, showing that the aggregation number significantly increased from ~40 for ODG to ~80 for DDDG. All new compounds were able to solubilize MPs from bacterial membranes, insect cells, as well as Madin Darby Canine Kidney (MDCK) cells. In particular, native human adenosine receptor (A2AR) and bacterial transporter BmrA were solubilized efficiently. A striking thermostability improvement of +13 and +9°C when ODG and DDG were respectively applied to wild type and full length A2AR. Taken together, this novel detergent series shows promising detergent potency for solubilization and stabilization of MPs and, thus, makes a valuable addition to the chemical toolbox available for extracting and handling these important but challenging membrane protein targets.
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