Hydrogenated Diglucose Detergents for Membrane-Protein Extraction and Stabilization

Abstract

We report herein the design and synthesis of a novel series of alkyl glycoside 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, respectively. Micellization in aqueous solution was studied by isothermal titration calorimetry, 1H NMR spectroscopy, and surface tensiometry. 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 to 9 mM for DDDG and ODG, respectively. Dynamic light scattering and analytical ultracentrifugation sedimentation velocity experiments were used to investigate the size and 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 membrane proteins (MPs) from bacterial membranes, insect cells, as well as the Madin–Darby canine kidney cells. In particular, native human adenosine receptor (A2AR) and bacterial transporter (BmrA) were solubilized efficiently. Striking thermostability improvements of +13 and +8 °C were observed 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 membrane proteins (MPs) and thus makes a valuable addition to the chemical toolbox available for extracting and handling these important but challenging MP targets.

Stabilization of Human Multidrug Resistance Protein 4 (MRP4/ABCC4) Using Novel Solubilization Agents

“…This was also observed for other membrane proteins using other detergents. 29 Further studies using other thermostability assays are required to explain the contribution of the chemical environment (lipid/detergent/polymer) and the membrane protein dynamics (in detergent or in SMA) on the shape of thermal shift curves.…”

Maltose-Based Fluorinated Surfactants for Membrane-Protein Extraction and Stabilization

“…Sugar-based surfactants are widely considered mild detergents and have been commonly used as solubilizing and non-denaturing detergents for MP applications including n-octyl-β-D-glucoside (OG), [2][3] n-decyl-β-D-maltoside (DM), 2 n-dodecyl-β-D-maltoside (DDM), [4][5][6] lactobionamides, 7 and thioglycoside such as n-octyl-β-D-thioglucopyranoside (OTG). [8][9] Recently, some sugar-based surfactants with branched polar headgroups or branched hydrophobic tails have been described for the same objective, namely, alkyl diglucosides (DigluM), 10 CALX-173-GK, 11 and laurylmaltose neopentylglycol (LMNG). [12][13][14] Fluorinated surfactants (FSs) bear several perfluorinated carbon atoms in their hydrophobic tail.…”

Cyclohexyl-α maltoside as a highly efficient tool for membrane protein studies

“…While several non-detergent systems for extraction or stabilization of MPs have been developed, e.g. , amphipols ( Popot, 2010 ), nanodiscs ( Denisov and Sligar, 2016 ), peptidiscs ( Carlson et al., 2018 ) or styrene maleic acid co-polymer lipid particles (SMALPs) ( Stroud et al., 2018 ), novel surfactants, including detergents and amphiphiles, are regularly reported ( Guillet et al., 2019 ; Chae et al., 2010 ; Breibeck and Rompel, 2019 ). Many of these new generation compounds are DDM analogues, building on its outstanding track record within MP research ( Chae et al., 2010 ; Breibeck and Rompel, 2019 ; Hong et al., 2010 ).…”